CRYSTALLINE FORMS OF (S)-7-([1,2,4]TRIAZOLO[1,5-a]PYRIDIN-6-YL)-4-(3,4-DICHLOROPHENYL)-1,2,3,4- TETRAHYDROISOQUINOLINE AND USE THEREOF

ABSTRACT

Novel [1,2,4]triazolo[1,5-a]pyridinyl-6-yl-substituted tetrahydroisoquinolines are described in the present invention. These compounds and crystalline forms SA1 and N-2 are used in the treatment of various neurological and physiological disorders. Methods of making these compounds and crystalline forms SA-1 and N-2 are also described in the present invention.

This application claims the benefit of U.S. Provisional PatentApplication Ser. No. 61/177,486, filed May 12, 2009, which is herebyincorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to compounds, crystalline forms,compositions, and methods for the treatment of various neurological andpsychological disorders, and the use of the compounds and crystallineforms in combination therapy. In particular, the present inventionrelates to such compounds, crystalline forms, compositions, and methods,where the compounds are novel[1,2,4]triazolo[1,5-a]pyridinyl-6-yl-substituted tetrahydroisoquinolinederivatives. Methods of making these compounds and crystalline formsSA-1 and N-2 are also described in the present invention.

BACKGROUND OF THE INVENTION

Monoamine reuptake inhibitors elevate extracellular levels of serotonin(5-HT), norepinephrine (NE) and/or dopamine (DA) in the brain by bindingto one or more of the transporters responsible for reuptake, namely theserotonin transporter (SERT), the norepinephrine transporter (NET) andthe dopamine transporter (DAT), thereby blocking reuptake of theneurotransmitter(s) from the synaptic cleft. Monoamine reuptakeinhibitors are an established drug class that has proven utility for thetreatment of a number of CNS disorders especially major depressivedisorder (MDD).

Since the introduction of tricylic antidepressants (TCAs) almost 50years ago, monoamine reuptake inhibitors with greatly improved safetyprofiles have significantly enhanced the treatment of depression.Although TCAs are very effective antidepressants, cardiovascular,anticholinergic and sedative side effects are common due to theinteraction of TCAs with muscarinic, histaminic and adrenergicreceptors. The revolutionary introduction of selective serotoninreuptake inhibitors (SSRIs) in the 1980s allowed a much larger patientpopulation to be treated because of the highly improved safety profile.Over the past decades, inhibitors that selectively block the reuptake ofNE or DA, or two of the three neurotransmitters simultaneously, havebecome available for the treatment of CNS disorders includingdepression, anxiety, obsessive compulsive disorder (OCD), attentiondeficit hyperactivity disorder (ADHD), pain and urinary incontinence.Two representative recent reviews (Liu and Molino, Annual Reports inMedicinal Chemistry, 42:13 (2007); Walter, Drug Dev. Res., 65:97 (2005))on monoamine reuptake inhibitors summarized the history and recentdevelopment in the monoamine reuptake inhibitor area.

Currently, the major effort in the field of monoamine reuptakeinhibitors is focused on improving antidepressant efficacy since 30-40%of patients do not respond to treatment with currently availableantidepressants. An additional major objective is to enhance the onsetof action. Current antidepressants typically require 2-6 weeks oftreatment before clinical efficacy is seen. Clinical trials exploringaugmentation strategies, in which a DA reuptake inhibitor or a dualNE/DA reuptake inhibitor is combined with an SSRI, have resulted inimproved efficacy in depressed patients refractory to SSRI treatmentalone (Patkar et. al, J. Clin. Psychopharmacol., 26:653 (2006); Zisooket al, Biol. Psychiat., 59:203 (2006)). The improved results fromclinical trials such as these serve to justify the considerable focus onthe development of inhibitors that simultaneously block the reuptake of5-HT, NE and DA. Because of the continued need for better drugs to treatdepression and the opportunities for new clinical indications, effortsto discover novel monoamine reuptake inhibitors continue unabated.

Methylphenidate, currently used for the treatment of attentiondeficit-hyperactivity disorder, is known to be selective for inhibitionof the DAT. Also, U.S. Pat. No. 5,444,070 discloses selective inhibitorsof dopamine reuptake as treatments for Parkinson's disease, drugaddiction or abuse including cocaine and amphetamines.

Selective norepinephrine reuptake inhibitors (NARI) have also beendisclosed. U.S. Pat. No. 6,352,986 describes methods of treatingattention deficit-hyperactivity disorder (ADHD), addictive disorders,and psychoactive substance use disorders with Reboxetine. Also,Atomoxetine (STRATTERA®) is currently marketed as a selective NETreuptake inhibitor for ADHD.

The use of selective serotonin reuptake inhibitors (SSRI) has been shownto be effective in treating depressive disorders. Sertraline,citalopram, escitalopram, paroxetine, fluoxetine and fluvoxamine arewell known examples of SSRIs used to treat disorders such as depression,obsessive compulsive disorder, and panic attacks. There are severalknown difficulties with the SSRI class of therapeutics, including theslow onset of action, unwanted side effects, and the existence of asignificant subset of the population that is not responsive to SSRItherapy. Recent effort in the clinical development of new SSRIs hasfocused on the treatment of premature ejaculation (PE) by takingadvantage of the ejaculation-delaying side effects of SSRIs. AlthoughSSRIs have been prescribed off-label to treat this condition, an SSRIwith rapid onset of action and rapid clearance could be preferred foron-demand treatment of PE. Dapoxetine (LY210448, 6), an SSRIstructurally related to fluoxetine with a shorter half-life, wasreported to be an effective and generally well tolerated treatment formen with moderate-to-severe PE in clinical trials (Feret, Formulary,40:227 (2005); Pryor et al, Lancet, 368:929 (2006)).

Selective inhibitors of DAT, NET, and SERT reuptake may also beco-administered with each other or with other drugs. U.S. Pat. No.5,532,244 discloses the use of serotonin reuptake inhibitors incombination with a serotonin 1A antagonist for the treatment ofobsessive-compulsive disorder, depression, and obesity. The use of aserotonin or norepinephrine reuptake inhibitor in combination with aneurokinin-1 receptor antagonist has been disclosed in U.S. Pat. No.6,121,261 for the treatment of ADHD. U.S. Pat. No. 4,843,071 disclosesthe use of a norepinephrine reuptake inhibitor in combination with anorepinephrine precursor in the treatment of obesity, drug abuse, ornarcolepsy. U.S. Pat. No. 6,596,741 discloses the use of a NE, DA, or5-HT inhibitor with either a neurokinin-1 receptor antagonist or aserotonin-1A antagonist for the treatment of a wide variety ofconditions.

Also advantageous is the use of compounds that inhibit one or more ofthe neurotransmitters at the same time. The antidepressant qualities ofthe dual NET and SERT reuptake inhibitor duloxetine is disclosed inEuropean Patent No. EP 273658. Venlafaxine is disclosed in U.S. Pat. No.4,535,186 as a reuptake inhibitor of both NE and 5-HT for the treatmentof depressive disorders. U.S. Pat. No. 6,635,675 discloses the use ofthe dual NE and 5-HT reuptake inhibitor milnacipran for the treatment ofchronic fatigue syndrome and fibromyalgia syndrome. In addition, dual NEand 5-HT reuptake inhibitors are also disclosed in U.S. Pat. No.6,136,083 for the treatment of depression. It is also recognized thatcompounds which inhibit the reuptake of NE, DA, and 5-HT in varyingratios not specifically mentioned here would also be advantageous.

As the first SNRI drug approved, venlafaxine has become one of thefirst-line choices for depression and anxiety disorder. An activemetabolite, desvenlafaxine, is also under clinical development for thetreatment of major depressive disorders. Preclinical studies alsoindicate that desvenlafaxine may be effective in relieving vasomotorsymptoms associated with menopause (e.g., hot flashes and night sweats)(Sorbera, et al, Drugs of Future., 31:304 (2006); Albertazzi, J. Br.Menopause Soc., 12:7 (2006)). Desvenlafaxine is reported to be inclinical development for the treatment of fibromyalgia and neuropathicpain, as well as vasomotor symptoms associated with menopause.

In addition to treating major depressive disorder, duloxetine wasapproved as the first agent for the treatment of painful diabeticneuropathy in the U.S. It also has been used for stress urinaryincontinence in women in Europe. In 2007, duloxetine was approved forthe treatment of generalized anxiety disorder in the U.S. Most recently,it was approved by the FDA for the management of fibromyalgia.

Milnacipran is currently available for use as an antidepressant inseveral countries outside the U.S. It is also under clinical developmentto assess its potential role in the treatment of fibromyalgia syndrome.

After more than a decade of use, bupropion, is considered a safe andeffective antidepressant, suitable for use as first-line treatment. Inaddition, it is approved for smoking cessation and seasonal affectivedisorder. It is also prescribed off-label to treat the sexualdysfunction induced by SSRIs. Bupropion is often referred to as anatypical antidepressant. It has much lower affinity for the monoaminetransporters compared with other monoamine reuptake inhibitors. Themechanism of action of bupropion is still uncertain but may be relatedto inhibition of dopamine and norepinephrine reuptake transporters as aresult of active metabolites. In a recently reported clinical trial,bupropion extended release (XL) had a sexual tolerability profilesignificantly better than that of escitalopram with similar remissionrates and Hospital Anxiety and Depression (HAD) total scores in patientswith major depressive disorder (Clayton et al. J. Clin. Psychiatry,67:736 (2006)).

Treating illnesses by inhibiting the reuptake of all three of themonoamines either through combination therapy or “triple inhibitors” mayhave clinical benefit as well. Triple inhibitors are considered to bethe next generation antidepressant (Liang and Richelson, PrimaryPsychiatry, 15(4):50 (2008)). Rationale for inclusion of a dopamineenhancing component in anti-depressant therapy includes observeddeficits in dopaminergic function, the success of combination therapywith dopamine agonists and traditional anti-depressants, and anincreased sensitivity in dopamine receptors due to chronicanti-depressant administration (Skolnick et al., Life Sciences,73:3175-3179 (2003)). Combination therapy with an SSRI and anoradrenaline and dopamine reuptake inhibitor was shown to be moreefficacious in patients with treatment-resistant depression (Lam et al,J. Clin. Psychiatry, 65(3):337-340 (2004)). Clinical studies using thecombination of bupropion and an SSRI or SNRI have showed improvedefficacy for the treatment of MDD in patients refractory to thetreatment with SSRIs, SNRIs, or bupropion alone (Zisook et al, Biol.Psychiat., 59:203 (2006); Papkostas, Depression and Anxiety, 23:178-181(2006); Trivedi et al, New Engl. J. Med., 354:1243 (2006)). Otherstudies using methylphenidate, both immediate release and extendedrelease formula, have shown it to be effective as an augmenting agent intreatment-resistant depression (Patkar et al, J. Clin. Psychopharmacol.,26:653 (2006); Masand et al, Depression and Anxiety, 7:89 (1998)). Inaddition, the combination of bupropion-SR with either SSRIs ornorepinephrine and dopamine reuptake inhibitors was found to induce lesssexual dysfunction than monotherapy (Kennedy et al, J. Clin. Psychiatry,63(3):181-186 (2002)). As such, inhibitory activity against DA reuptake,in addition to NE and 5-HT reuptake, is expected to provide a more rapidonset of anti-depressant effect than other mixed inhibitors which areselective for NET and SERT over DAT. PCT International Publication Nos.WO 03/101453 and WO 97/30997 disclose a class of compounds which areactive against all three monoamine transporters. Also, PCT InternationalPatent Publication No. WO 03/049736 discloses a series of 4-substitutedpiperidines, each of which displays similar activity against DA, NE, and5-HT transporters. Bicyclo[2.2.1]Heptanes (Axford et al., Bioorg. Med.Chem. Lett., 13:3277-3280 (2003)) and azabicyclo[3.1.0]Hexanes (Skolnicket al., Eur. J. Pharm., 461:99-104 (2003)) are also described as tripleinhibitors of the three monoamine transporters.1-(3,4-Dichlorophenyl)-3-azabicyclo[3.1.0]Hexane has been shown to beefficacious in treating depression in clinical trials (Beer et al, J.Clin. Pharmacol., 44:1360-1367 (2004)). Current widely used anti-obesitydrug sibutramine is believed to work through the inhibition of all threetransporters DAT, SERT, and NET (Ryan, Pharmacotherapy of Obesity,245-266 (2004)).

Recent drug approvals with SNRIs for treatment of fibromyalgia anddiabetic neuropathy reinforce the utility of this drug class in thetreatment of neuropathic pain. Other largely untapped areas which remainto be exploited with this drug class include sexual dysfunction, such aspremature ejaculation, irritable bowel syndrome, obesity,neurodegenerative diseases such as Parkinson's disease, restless legsyndrome, and substance abuse and addiction.

There is still a large need for compounds that block the reuptake ofnorepinephrine, dopamine, and serotonin and treat various neurologicaland psychological disorders.

The present invention is directed achieving this objective.

SUMMARY OF THE INVENTION

The present invention relates to crystalline Form SA-1 of

wherein the carbon atom designated * is in the S configuration, or apharmaceutically acceptable salt thereof or a solvate thereof.

The present invention also relates to crystalline Form N-2 of

wherein the carbon atom designated * is in the S configuration, or apharmaceutically acceptable salt thereof or a solvate thereof.

Another aspect of the present invention relates to a process forpreparation of a product compound of Formula (I)

wherein the carbon atom designated * is in the R or S configuration.This process comprises treating a first intermediate compound of Formula(II):

with an acid under conditions effective to produce the product compound.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates experimental and simulated powder X-ray diffraction(PXRD) patterns (CuKα λ=1.54178 Å at T=room temperature) of Form SA-1.

FIG. 2 illustrates the differential scanning calorimetry (DSC) patternof Form SA-1.

FIG. 3 illustrates thermogravimetric analysis (TGA) of Form SA-1.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to a compound of formula (I):

wherein:

-   the carbon atom designated * is in the R or S configuration;-   or a pharmaceutically acceptable salt thereof or a solvate thereof.

As used above, and throughout the description of the invention, thefollowing terms, unless otherwise indicated, shall be understood to havethe following meanings:

The term “compounds of the invention”, and equivalent expressions, aremeant to embrace compounds of general formula (I) as hereinbeforedescribed, which expression includes the pharmaceutically acceptablesalts and the solvates, e.g. hydrates, where the context so permits.Similarly, reference to intermediates, whether or not they themselvesare claimed, is meant to embrace their salts, and solvates, where thecontext so permits. For the sake of clarity, particular instances whenthe context so permits are sometimes indicated in the text, but theseinstances are purely illustrative and it is not intended to excludeother instances when the context so permits.

The term “pharmaceutically acceptable salts” means the relativelynon-toxic, inorganic, and organic acid addition salts, and base additionsalts, of compounds of the present invention. These salts can beprepared in situ during the final isolation and purification of thecompounds. In particular, acid addition salts can be prepared byseparately reacting the purified compound in its free base form with asuitable organic or inorganic acid and isolating the salt thus formed.Exemplary acid addition salts include the hydrobromide, hydrochloride,sulfate, bisulfate, phosphate, nitrate, acetate, oxalate, valerate,oleate, palmitate, stearate, laurate, borate, benzoate, lactate,phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate,naphthylate, mesylate, glucoheptonate, lactiobionate, sulphamates,malonates, salicylates, propionates, methylene-bis-b-hydroxynaphthoates,gentisates, isethionates, di-p-toluoyltartrates, methane-sulphonates,ethanesulphonates, benzenesulphonates, p-toluenesulphonates,cyclohexylsulphamates and quinateslaurylsulphonate salts, and the like(see, for example, Berge et al., “Pharmaceutical Salts,” J. Pharm. Sci.,66:1-9 (1977) and Remington's Pharmaceutical Sciences, 17th ed., MackPublishing Company, Easton, Pa., 1985, p. 1418, which are herebyincorporated by reference in their entirety). Base addition salts canalso be prepared by separately reacting the purified compound in itsacid form with a suitable organic or inorganic base and isolating thesalt thus formed. Base addition salts include pharmaceuticallyacceptable metal and amine salts. Suitable metal salts include thesodium, potassium, calcium, barium, zinc, magnesium, and aluminum salts.The sodium and potassium salts are preferred. Suitable inorganic baseaddition salts are prepared from metal bases which include, for example,sodium hydride, sodium hydroxide, potassium hydroxide, calciumhydroxide, aluminium hydroxide, lithium hydroxide, magnesium hydroxide,and zinc hydroxide. Suitable amine base addition salts are prepared fromamines which have sufficient basicity to form a stable salt, andpreferably include those amines which are frequently used in medicinalchemistry because of their low toxicity and acceptability for medicaluse, such as ammonia, ethylenediamine, N-methyl-glucamine, lysine,arginine, ornithine, choline, N,N′-dibenzylethylenediamine,chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine,diethylamine, piperazine, tris(hydroxymethyl)-aminomethane,tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine,dehydroabietylamine, N-ethylpiperidine, benzylamine,tetramethylammonium, tetraethylammonium, methylamine, dimethylamine,trimethylamine, ethylamine, basic amino acids, e.g., lysine andarginine, dicyclohexylamine, and the like.

The term “substantially pure” refers to chemical purity and form purity.For example, substantially pure Form SA-1 (or Form N-2) comprises atleast about 95 wt %, preferably at least about 98 wt %, more preferablyat least about 99 wt % of Form SA-1 and less than about 5 wt %,preferably less than about 2 wt %, and more preferably less than about 1wt % of other compounds having a different chemical structure than theS-enantiomer of Formula (I). Additionally, substantially pure Form SA-1(or Form N-2) comprises at least about 95 wt %, preferably at leastabout 98 wt %, more preferably at least about 99 wt % of Form SA-1 andless than about 5 wt %, preferably less than about 2 wt %, and morepreferably less than about 1 wt % of any other crystalline form of theS-enantiomer of Formula (I). This means that the Form SA-1 (or Form N-2)preferably contains less than about 5 wt % of other compounds, and lessthan about 5 wt % of any other form (also referred to as “phasehomogenicity”).

The term “therapeutically effective amounts” is meant to describe anamount of compound of the present invention effective in increasing thelevels of serotonin, norepinephrine, or dopamine at the synapse and thusproducing the desired therapeutic effect. Such amounts generally varyaccording to a number of factors well within the purview of ordinarilyskilled artisans given the description provided herein to determine andaccount for. These include, without limitation: the particular subject,as well as its age, weight, height, general physical condition, andmedical history, the particular compound used, as well as the carrier inwhich it is formulated and the route of administration selected for it;and, the nature and severity of the condition being treated.

The term “pharmaceutical composition” means a composition comprising acompound of formula (I) and at least one component comprisingpharmaceutically acceptable carriers, diluents, adjuvants, excipients,or vehicles, such as preserving agents, fillers, disintegrating agents,wetting agents, emulsifying agents, suspending agents, sweeteningagents, flavoring agents, perfuming agents, antibacterial agents,antifungal agents, lubricating agents and dispensing agents, dependingon the nature of the mode of administration and dosage forms. Examplesof suspending agents include ethoxylated isostearyl alcohols,polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminum metahydroxide, bentonite, agar—agar and tragacanth,or mixtures of these substances. Prevention of the action ofmicroorganisms can be ensured by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, sorbic acid, andthe like. It may also be desirable to include isotonic agents, forexample sugars, sodium chloride, and the like. Prolonged absorption ofthe injectable pharmaceutical form can be brought about by the use ofagents delaying absorption, for example, aluminum monosterate andgelatin. Examples of suitable carriers, diluents, solvents, or vehiclesinclude water, ethanol, polyols, suitable mixtures thereof, vegetableoils (such as olive oil), and injectable organic esters such as ethyloleate. Examples of excipients include lactose, milk sugar, sodiumcitrate, calcium carbonate, and dicalcium phosphate. Examples ofdisintegrating agents include starch, alginic acids, and certain complexsilicates. Examples of lubricants include magnesium stearate, sodiumlauryl sulphate, talc, as well as high molecular weight polyethyleneglycols.

The term “pharmaceutically acceptable” means it is, within the scope ofsound medical judgment, suitable for use in contact with the cells ofhumans and lower animals without undue toxicity, irritation, allergicresponse and the like, and are commensurate with a reasonablebenefit/risk ratio.

The term “pharmaceutically acceptable dosage forms” means dosage formsof the compound of the invention, and includes, for example, tablets,dragees, powders, elixirs, syrups, liquid preparations, includingsuspensions, sprays, inhalants tablets, lozenges, emulsions, solutions,granules, capsules, and suppositories, as well as liquid preparationsfor injections, including liposome preparations. Techniques andformulations generally may be found in Remington's PharmaceuticalSciences, Mack Publishing Co., Easton, Pa., latest edition.

In one preferred embodiment of the present invention, the compound offormula (I) is a (+)-stereoisomer.

In another preferred embodiment of the present invention, the compoundof formula (I) is a (−)-stereoisomer.

Another more preferred embodiment of the present invention is thecompound of formula (I) wherein the carbon atom designated * is in the Rconfiguration.

Another more preferred embodiment of the present invention is thecompound of formula (I) wherein the carbon atom designated * is in the Sconfiguration.

In another preferred embodiment of the present invention, the compoundof formula (I) is a (S)(+)-stereoisomer.

In yet another preferred embodiment of the present invention, thecompound of formula (I) is a (R)(−)-stereoisomer.

Another preferred embodiment of the present invention is a mixture ofstereoisomeric compounds of formula (I) wherein * is in the S or Rconfiguration.

Single enantiomers, any mixture of enantiomers, including racemicmixtures, or diastereomers (both separated and as any mixtures) of thecompounds of the present invention are also included within the scope ofthe invention.

The scope of the present invention also encompasses active metabolitesof the present compounds.

The present invention also includes compounds of formula (I), whereinone or more of the atoms, e.g., C or H, are replaced by thecorresponding radioactive isotopes of that atom (e.g., C replaced by ¹⁴Cand H replaced by ³H), or a stable isotope of that atom (e.g., Creplaced by ¹³C or H replaced by ²H). Such compounds have a variety ofpotential uses, e.g., as standards and reagents in determining theability of a potential pharmaceutical to bind to neurotransmitterproteins. In addition, in the case of stable isotopes, such compoundsmay have the potential to favorably modify the biological properties,e.g., pharmacological and/or pharmacokinetic properties, of compounds offormula (I). The details concerning selection of suitable sites forincorporating radioactive isotopes into the compounds are known to thoseskilled in the art.

Another aspect of the present invention relates to a crystalline form of7-([1,2,4]triazolo[1,5-a]pyridinyl-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline,in particular, Form SA-1 or Form N-2, as described herein. For purposesof clarification, the free base racemate ofrac-7-([1,2,4]triazolo[1,5-a]pyridinyl-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolineis represented by Formula (I). Forms SA-1 and N-2 are particularcrystalline forms of the S-enantiomer of Formula (I)((S)-7-([1,2,4]triazolo[1,5-a]pyridinyl-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline),as described herein.

Thus, one embodiment of the present invention relates to Form SA-1. Oneaspect of this embodiment of the present invention relates to Form SA-1,characterized by the following unit cell parameters:

Cell Dimensions:

-   -   a=11.0668(9) Å    -   b=7.3750(6) Å    -   c=15.3927(14) Å

alpha=90°

-   -   beta=100.594(7)°    -   gamma=90°

Space group: Monoclinic, P2₁

Volume: 1234.90(18) Å³

Z, Calculated Density: 2, 1.363 Mg/m³

Another aspect of this embodiment of the present invention relates toForm SA-1, characterized by fractional atomic coordinates within theunit cell as listed in Table 6, Atomic Coordinates.

A further aspect of this embodiment of the present invention relates toForm SA-1 with characteristic peaks in the powder X-ray diffractionpattern at values of 2 theta of 5.8±0.1, 8.1±0.1, 9.1±0.1, 10.8±0.1,11.7±0.1, 13.0±0.1, 13.3±0.1, 14.5±0.1, 15.1±0.1, 15.4±0.1, 16.2±0.1,and 16.8±0.1, at a temperature between about 20° C. and about 25° C.,based on a high quality pattern collected with a diffractometer (cuKαe)with a spinning capillary with 2θ calibrated with a National Instituteof Standards and Technology (NIST) or other suitable standard.

Another aspect of this embodiment of the present invention relates toForm SA-1 characterized by a melt with decomposition endotherm withonset typically of 85° C.

A further aspect of this embodiment of the present invention relates tosubstantially pure Form SA-1.

Another embodiment of the present invention relates to Form N-2. Oneaspect of this embodiment of the present invention relates to Form N-2,characterized by the following unit cell parameters:

Cell Dimensions:

-   -   a=7.1183(2) Å    -   b=21.2160(7) Å    -   c=26.3602(9) Å    -   alpha=90°    -   beta=90°    -   gamma=90°

Space group: Orthorhombic, P2₁2₁2₁

Volume: 3981.0(2) Å³

Z, Calculated Density: 8, 1.441 Mg/m³

Another aspect of this embodiment of the present invention relates toForm N-2, characterized by fractional atomic coordinates within the unitcell as listed in Table 8, Atomic Coordinates.

A further aspect of this embodiment of the present invention relates toForm N-2 with characteristic peaks in the powder X-ray diffractionpattern at values of 2 theta of 8.3±0.1, 8.9±0.1, 10.9±0.1, 14.2±0.1,14.7±0.1, 16.7±0.1, 17.3±0.1, 18.0±0.1, 18.4±0.1, 18.8±0.1, 20.2±0.1,and 21.9±0.1, at a temperature between about 20° C. and about 25° C.,based on a high quality pattern collected with a diffractometer (cuKα)with a spinning capillary with 20 calibrated with a NIST or othersuitable standard.

Another aspect of this embodiment of the present invention relates toForm N-2 characterized by a melt with decomposition endotherm with onsettypically of about 250° C.

A further aspect of this embodiment of the present invention relates tosubstantially pure Form N-2.

Another aspect of the present invention is a pharmaceutical compositioncontaining a therapeutically effective amount of the compound of formula(I) or a crystalline form as described herein and a pharmaceuticallyacceptable carrier.

Another aspect of the present invention relates to a method of treatinga disorder which is created by or is dependent upon decreasedavailability of serotonin, norepinephrine, or dopamine. The methodinvolves administering to a patient in need of such treatment atherapeutically effective amount of a compound of formula (I), acrystalline form of a compound of formula (I), or a pharmaceuticallyacceptable salt thereof. The method of the present invention is capableof treating subjects afflicted with various neurological and psychiatricdisorders including, without limitation: attention deficit hyperactivitydisorder (ADHD), cognition impairment, anxiety disorders, generalizedanxiety disorder (GAD), panic disorder, bipolar disorder or manicdepression or manic-depressive disorder, obsessive compulsive disorder(OCD), posttraumatic stress disorder (PTSD), acute stress disorder,social phobia, simple phobias, pre-menstrual dysphoric disorder (PMDD),social anxiety disorder (SAD), major depressive disorder (MDD),postnatal depression, dysthymia, depression associated with Alzheimer'sdisease, Parkinson's disease, or psychosis, supranuclear palsy, eatingdisorders, obesity, anorexia nervosa, bulimia nervosa, binge eatingdisorder, diabetes, ischemic diseases, pain, substance abuse disorders,chemical dependencies, nicotine addiction, cocaine addiction,amphetamine addiction, alcohol addiction, Lesch-Nyhan syndrome,neurodegenerative diseases, Parkinson's disease, late luteal phasesyndrome or narcolepsy, psychiatric symptoms, anger, rejectionsensitivity, movement disorders, extrapyramidal syndrome, Tic disorders,restless leg syndrome (RLS), tardive dyskinesia, supranuclear palsy,sleep related eating disorder (SRED), night eating syndrome (NES),stress urinary incontinence (SUI), migraine, neuropathic pain, diabeticneuropathy, lower back pain, fibromyalgia syndrome (FS), osteoarthritispain, arthritis pain, chronic fatigue syndrome (CFS), sexualdysfunction, premature ejaculation, male impotence, thermoregulatorydisorders (e.g., hot flashes associated with menopause), and irritablebowel syndrome (IBS).

The compounds/crystalline forms provided herein are particularly usefulin the treatment of these and other disorders due, at least in part, totheir ability to selectively bind to the transporter proteins forcertain neurochemicals with a greater affinity than to the transporterproteins for other neurochemicals.

In another embodiment of the present invention, the above method furtherinvolves administering a therapeutically effective amount of a serotonin1A receptor antagonist or a pharmaceutically acceptable salt thereof.Suitable serotonin 1A receptor antagonists include WAY 100135 andspiperone. WAY 100135(N-(t-butyl)-3-[a-(2-methoxyphenyl)piperazin-1-yl]-2 phenylpropanamide)is disclosed as having an affinity for the serotonin 1A receptor in U.S.Pat. No. 4,988,814 to Abou-Gharbia et al., which is hereby incorporatedby reference in its entirety. Also, Cliffe et al., J Med Chem 36:1509-10(1993), which is hereby incorporated by reference in its entirety,showed that the compound is a serotonin 1A antagonist. Spiperone(844-(4-fluorophenyl)-4-oxobutyl]-1-phenyl-1,3,8-triazaspiro[4,5]decan-4-one)is a well-known compound and is disclosed in U.S. Pat. Nos. 3,155,669and 3,155,670, which are hereby incorporated by reference in theirentirety. The activity of spiperone as a serotonin 1A antagonist isdescribed in Middlemiss et al., Neurosc and Biobehav Rev. 16:75-82(1992), which is hereby incorporated by reference in its entirety.

In another embodiment of the present invention, the above method furtherinvolves administering a therapeutically effective amount of a selectiveneurokinin-1 receptor antagonist or pharmaceutically acceptable saltthereof. Neurokinin-1 receptor antagonists that can be used incombination with the compound of formula (I) or crystalline form, in thepresent invention are fully described, for example, in U.S. Pat. Nos.5,373,003, 5,387,595, 5,459,270, 5,494,926, 5,162,339, 5,232,929,5,242,930, 5,496,833, and 5,637,699; PCT International PatentPublication Nos. WO 90/05525, 90/05729, 94/02461, 94/02595, 94/03429,94/03445, 94/04494, 94/04496, 94/05625, 94/07843, 94/08997, 94/10165,94/10167, 94/10168, 94/10170, 94/11368, 94/13639, 94/13663, 94/14767,94/15903, 94/19320, 94/19323, 94/20500, 91/09844, 91/18899, 92/01688,92/06079, 92/12151, 92/15585, 92/17449, 92/20661, 92/20676, 92/21677,92/22569, 93/00330, 93/00331, 93/01159, 93/01165, 93/01169, 93/01170,93/06099, 93/09116, 93/10073, 93/14084, 93/14113, 93/18023, 93/19064,93/21155, 93/21181, 93/23380, 93/24465, 94/00440, 94/01402, 94/26735,94/26740, 94/29309, 95/02595, 95/04040, 95/04042, 95/06645, 95/07886,95/07908, 95/08549, 95/11880, 95/14017, 95/15311, 95/16679, 95/17382,95/18124, 95/18129, 95/19344, 95/20575, 95/21819, 95/22525, 95/23798,95/26338, 95/28418, 95/30674, 95/30687, 95/33744, 96/05181, 96/05193,96/05203, 96/06094, 96/07649, 96/10562, 96/16939, 96/18643, 96/20197,96/21661, 96/29304, 96/29317, 96/29326, 96/29328, 96/31214, 96/32385,96/37489, 97/01553, 97/01554, 97/03066, 97/08144, 97/14671, 97/17362,97/18206, 97/19084, 97/19942, 97/21702, and 97/49710; and in U.K. PatentApplication Nos. 2 266 529, 2 268 931, 2 269 170, 2 269 590, 2 271 774,2 292 144, 2 293168, 2 293 169, and 2 302 689; European PatentPublication Nos. EP 0 360 390, 0 517 589, 0 520 555, 0 522 808, 0 528495, 0 532 456, 0 533 280, 0 536 817, 0 545 478, 0 558 156, 0 577 394, 0585 913, 0 590 152, 0 599 538, 0 610 793, 0 634 402, 0 686 629, 0 693489, 0 694 535, 0 699 655, 0 394 989, 0 428 434, 0 429 366, 0 430 771, 0436 334, 0 443 132, 0 482 539, 0 498 069, 0 499 313, 0 512 901, 0 512902, 0 514 273, 0 514 274, 0 514 275, 0 514 276, 0 515 681, 0 699 674, 0707 006, 0 708 101, 0 709 375, 0 709 376, 0 714 891, 0 723 959, 0 733632 and 0 776 893, which are hereby incorporated by reference in theirentirety. The preparations of such compounds are fully described in theaforementioned patents and publications.

In another embodiment of the present invention, the above method furtherinvolves administering a therapeutically effective amount of anorepinephrine precursor or a pharmaceutically acceptable salt thereof.Suitable norepinephrine precursors include L-tyrosine andL-phenylalanine.

Another embodiment of the present invention is a method of inhibitingsynaptic norepinephrine uptake in a patient in need thereof. The methodinvolves administering a therapeutically effective inhibitory amount ofa compound of formula (I) or a crystalline form as described herein.

Another embodiment of the present invention is a method of inhibitingsynaptic serotonin uptake in a patient in need thereof. The methodinvolves administering a therapeutically effective inhibitory amount ofa compound of formula (I) or a crystalline form as described herein.

Another embodiment of the present invention is a method of inhibitingsynaptic dopamine uptake in a patient in need thereof. The methodinvolves administering a therapeutically effective inhibitory amount ofa compound of formula (I) or a crystalline form as described herein.

Another embodiment of the present invention is a therapeutic methoddescribed herein, where the (+)-stereoisomer of the compound of formula(I) is employed.

Another embodiment of the present invention is a therapeutic methoddescribed herein, where the (−)-stereoisomer of the compound of formula(I) is employed.

Another embodiment of the present invention is a kit comprising acompound of formula (I) or a crystalline form as described herein, andat least one compound selected from the group consisting of: a serotonin1A receptor antagonist compound, a selective neurokinin-1 receptorantagonist compound, and a norepinephrine precursor compound.

Another embodiment of the present invention relates to a method oftreating a disorder referred to in the above-mentioned embodiments in apatient in need thereof. The method involves inhibiting synapticnorepinephrine, dopamine, and serotonin uptake by administering atherapeutically effective inhibitory amount of the compound of formula(I) or a crystalline form as described herein, which functions as atriple acting norepinephrine, dopamine, and serotonin uptake inhibitor.

Another embodiment of the present invention relates to a method forinhibiting serotonin uptake in mammals. The method involvesadministering to a mammal requiring increased neurotransmission ofserotonin a pharmaceutically effective amount of the compound of formula(I) or a crystalline form as described herein.

Another embodiment of the present invention relates to a method forinhibiting dopamine uptake in mammals. The method involves administeringto a mammal requiring increased neurotransmission of dopamine apharmaceutically effective amount of the compound of formula (I) or acrystalline form as described herein.

Another embodiment of the present invention relates to a method forinhibiting norepinephrine uptake in mammals. The method involvesadministering to a mammal requiring increased neurotransmission ofnorepinephrine a pharmaceutically effective amount of the compound offormula (I) or a crystalline form as described herein.

Another embodiment of the present invention relates to a method ofsuppressing the desire of humans to smoke. The method involvesadministering to a human in need of such suppression an effective dose,to relieve the desire to smoke, of the compound of formula (I) or acrystalline form as described herein.

Another embodiment of the present invention relates to a method ofsuppressing the desire of humans to consume alcohol. The method involvesadministering to a human in need of such suppression an effective dose,to relieve the desire to consume alcohol, of the compound of formula (I)or a crystalline form as described herein.

Another embodiment of the present invention relates to a process forpreparation of a product compound of Formula (I). This process comprisestreating a first intermediate compound of Formula (II):

with an acid under conditions effective to produce the product compound.

Suitable acids include, but are not limited to, sulfuric acid,methansulfonic acid, phosphoric acid, and L-tartaric acid.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination.

Compounds according to the invention, for example, starting materials,intermediates, or products, are prepared as described herein or by theapplication or adaptation of known methods, by which is meant methodsused heretofore or described in the literature.

Compounds useful according to the invention may be prepared by theapplication or adaptation of known methods, by which is meant methodsused heretofore or described in the literature, for example, thosedescribed by Larock, Comprehensive Organic Transformations, Wiley-VCHpublishers, New York (1989), which is hereby incorporated by referencein its entirety.

A compound of formula (I) including a group containing one or morenitrogen ring atoms, may be converted to the corresponding compoundwherein one or more nitrogen ring atom of the group is oxidized to anN-oxide, preferably by reacting with a peracid, for example peraceticacid in acetic acid or m-chloroperoxybenzoic acid in an inert solventsuch as dichloromethane, at a temperature from about room temperature toreflux, preferably at elevated temperature.

In the reactions described hereinafter, it may be necessary to protectreactive functional groups, for example hydroxy, amino, imino, thio, orcarboxy groups, where these are desired in the final product, to avoidtheir unwanted participation in the reactions. Conventional protectinggroups may be used in accordance with standard practice (e.g., Wuts etal., Protective Groups in Organic Chemistry (4^(th) Edition), Wiley(2006), and McOmie, Protective Groups in Organic Chemistry, Plenum Press(1973), which are hereby incorporated by reference in their entirety).

The novel tetrahydroisoquinoline reuptake inhibitors of Formula (I) ofthis invention can be prepared by the synthetic route depicted in Scheme1.

1-(3-Methoxyphenyl)-N-methylmethanamine is reacted with3,4-dichlorophenacyl bromide in the presence of triethylamine to give1-(3,4-dichlorophenyl)-2-((3-methoxybenzyl)(methyl)amino)ethanone.Reduction of this ketone by sodium borohydride yields1-(3,4-dichlorophenyl)-2-(3-methoxybenzyl)(methyl)amino)ethanol, whichundergoes acid mediated cyclization to give4-(3,4-dichlorophenyl)-7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline.This racemic tetraisoquinoline derivative can be separated via chiralHPLC or supercritical fluid chromatography (SFC) to give the singleenantiomers. Alternatively, the chiral separation can be achieved byrecrystallization using chiral acids such as di-p-toluoyl-D-tartaricacid or di-p-toluoyl-L-tartaric acid.

The4-(3,4-dichlorophenyl)-7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolineis converted to the correspondent phenol by the treatment with 48% HBrunder reflux. The resulting phenol is then converted the correspondenttriflate which is further transformed to the correspondent pinacolborate derivative4-(3,4-dichlorophenyl)-2-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinoline.Demethylation using 1-chloroethyl chloroformate followed by Bocprotection gives tert-butyl4-(3,4-dichlorophenyl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate.Reaction of this Boc protected tetraisoquinoline with6-bromo-[1,2,4]triazolo[1,5-a]pyridine under Suzuki coupling conditionsaffords tert-butyl7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate,which is then deprotected by TFA to give7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolineor formula (I).

An alternative synthetic route of preparing compounds of Formula (I) inthis invention is depicted in Scheme 2.

Suzuki coupling of 3-formylphenylboronic acid and6-bromo-[1,2,4]triazolo[1,5-a]pyridine gives3-([1,2,4]triazolo[1,5-a]pyridin-6-yl)benzaldehyde. This aldehydeundergoes a reductive amination to give2-(3-([1,2,4]triazolo[1,5-a]pyridin-6-yl)benzylamino)-1-(3,4-dichlorophenyl)ethanol,which is then subjected to sulfuric acid mediated cyclization to provide7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline.

A synthetic route of preparing L-tartrate salts of the present inventionis depicted in Scheme 3.

Compounds of formula (I) may be obtained in enantiomerically enriched(R) and (S) form by crystallization with chiral salts as well known toone skilled in the art, or alternatively, may be isolated through chiralHPLC employing commercially available chiral columns.

It will be appreciated that compounds according to the present inventionmay contain asymmetric centers. These asymmetric centers mayindependently be in either the R or S configuration and such compoundsare able to rotate a plane of polarized light in a polarimeter. If saidplane of polarized light is caused by the compound to rotate in acounterclockwise direction, the compound is said to be the (−)stereoisomer of the compound. If said plane of polarized light is causedby the compound to rotate in a clockwise direction, the compound is saidto be the (+) stereoisomer of the compound. It will be apparent to thoseskilled in the art that certain compounds useful according to theinvention may also exhibit geometrical isomerism. It is to be understoodthat the present invention includes individual geometrical isomers andstereoisomers and mixtures thereof, including racemic mixtures, ofcompounds of formula (I) hereinabove. Such isomers can be separated fromtheir mixtures, by the application or adaptation of known methods, forexample chromatographic techniques and recrystallization techniques, orthey are separately prepared from the appropriate isomers of theirintermediates.

Radiolabelled compounds of the invention are synthesized by a number oftechniques well known to those of ordinary skill in the art, e.g., byusing starting materials incorporating therein one or moreradioisotopes. Compounds of the present invention where a stableradioisotope, such as carbon-14, tritium, iodine-121, or anotherradioisotope, has been introduced synthetically are useful diagnosticagents for identifying areas of the brain or central nervous system thatmay be affected by disorders where norepinephrine, dopamine, orserotonin transporters and their uptake mechanism are implicated.

Crystalline forms may be prepared by a variety of methods, including forexample, crystallization or recrystallization from a suitable solvent,sublimation, growth from a melt, solid state transformation from anotherphase, crystallization from a supercritical fluid, and jet spraying.Techniques for crystallization or recrystallization of crystalline formsfrom a solvent mixture include, for example, evaporation of the solvent,decreasing the temperature of the solvent mixture, crystal seeding asupersaturated solvent mixture of the molecule and/or salt, freezedrying the solvent mixture, and addition of antisolvents(countersolvents) to the solvent mixture. High throughputcrystallization techniques may be employed to prepare crystalline formsincluding polymorphs. Crystals of drugs, including polymorphs, methodsof preparation, and characterization of drug crystals are discussed inBryn et al., Solid-State Chemistry of Drugs, 2^(nd) Edition, SSCI, WestLafayette, Ind. (1999), which is hereby incorporated by reference in itsentirety.

For crystallization techniques that employ solvent, the choice ofsolvent or solvents is typically dependent upon one or more factors,such as solubility of the compound, crystallization technique, and vaporpressure of the solvent, or the ability to afford a substantially purecrystalline form. Combinations of solvents may be employed, for example,the compound may be solubilized into a first solvent to afford asolution, followed by the addition of an antisolvent to decrease thesolubility of the compound in the solution and to afford the formationof crystals. An antisolvent is a solvent in which the compound has lowsolubility.

In one method to prepare crystals, a compound is suspended and/orstirred in a suitable solvent to afford a slurry, which may be heated topromote complete or partial dissolution. The term “slurry”, as usedherein, means a saturated solution of the compound, which may alsocontain an additional amount of the compound to afford a heterogeneousmixture of the compound and a solvent at a given temperature.

Seed crystals may be added to any crystallization mixture to promotecrystallization. Seeding may be employed to control growth of aparticular polymorph or to control the particle size distribution of thecrystalline product and/or afford a substantially pure crystalline form.Accordingly, calculation of the amount of seeds needed depends on thesize of the seed available and the desired size of an average productparticle as described, for example, in Mullin et al., “ProgrammedCooling of Batch Crystallizers,” Chemical Engineering Science,26:369-377 (1971), which is hereby incorporated by reference in itsentirety,. In general, seeds of small size are needed to controleffectively the growth of crystals in the batch. Seed of small size maybe generated by sieving, milling, or micronizing of large crystals, orby micro-crystallization of solutions. Care should be taken that millingor micronizing of crystals does not result in any change incrystallinity of the desired crystal form (i.e., change to amorphous orto another polymorph).

A cooled crystallization mixture may be filtered under vacuum, and theisolated solids may be washed with a suitable solvent, such as coldrecrystallization solvent, and dried under a nitrogen purge to affordthe desired crystalline form. The isolated solids may be analyzed by asuitable spectroscopic or analytical technique, such as solid statenuclear magnetic resonance, X-Ray powder diffraction, or the like, toassure formation of the preferred crystalline form of the product. Theresulting crystalline form is typically produced in an amount of greaterthan about 70 weight percent isolated yield, preferably greater than 90weight percent isolated yield, based on the weight of the compoundoriginally employed in the crystallization procedure. The product may beco-milled or passed through a mesh screen to delump the product, ifnecessary.

Crystalline forms may be prepared, for example, directly from thereaction medium of the process for preparing a compound of formula (I).This may be achieved, for example, by employing in the final processstep a solvent or a mixture of solvents from which Form SA-1 or Form N-2may be crystallized. Alternatively, crystalline forms may be obtained bydistillation or solvent addition techniques. Suitable solvents for thispurpose include, for example, non-polar solvents and polar solvents,including protic polar solvents such as alcohols, and aprotic polarsolvents such as ketones, the details and selection of which are knownto those skilled in the art.

The presence of more than one polymorph in a sample may be determined bytechniques such as powder X-Ray diffraction (PXRD) or solid statenuclear magnetic resonance spectroscopy (SSNMR). For example, thepresence of extra peaks in an experimentally measured PXRD pattern whencompared with a simulated PXRD pattern may indicate more than onepolymorph in the sample. The simulated PXRD may be calculated fromsingle crystal X-Ray data (see Smith, “A FORTRAN Program for CalculatingX-Ray Powder Diffraction Patterns,” Lawrence Radiation Laboratory,Livermore, Calif., UCRL-7196 (April 1963), which is hereby incorporatedby reference in its entirety). In one aspect, Form SA-1 or Form N-2Hasphase homogeneity indicated by less than 5 percent, preferably less than2 percent, and more preferably less than 1 percent of the total peakarea in the experimentally measured PXRD pattern arising from the extrapeaks that are absent from a simulated PXRD pattern.

Preferably, the crystallization technique provides a product comprisingsubstantially pure Form SA-1 or Form N-2. The crystallized materialpreferably comprises at least 95 wt % of Form SA-1/Form N-1, based onthe weight of the compound of formula (I) in the composition. Theremaining material may comprise other form(s) of the compound and/orreaction impurities and/or processing impurities arising from itspreparation. The presence of reaction impurities and/or processingimpurities may be determined by analytical techniques known in the art,such as, for example, chromatography, nuclear magnetic resonancespectroscopy, mass spectrometry, or infrared spectroscopy.

Form SA-1 and Form N-1 can be characterized using various techniques,which are well known to those of ordinary skill in the art. Examples ofcharacterization methods include, but are not limited to, single crystalX-Ray diffraction, powder X-Ray diffraction (PXRD), simulated powderX-Ray patterns (Yin et al., American Pharmaceutical Review, 6(2):80(2003), which is hereby incorporated by reference in its entirety),differential scanning calorimetry (DSC), solid-state ¹³C NMR (Earl etal., J. Magn. Reson., 48:35-54 (1982), which is hereby incorporated byreference in its entirety), Raman spectroscopy, infrared spectroscopy,moisture sorption isotherms, thermogravimetric analysis (TGA), and hotstage techniques.

The forms may be characterized and distinguished using single crystalX-ray diffraction, which is based on unit cell measurements of a singlecrystal of Form SA-1 or Form N-2. A detailed description of unit cellsis provided in Stout et al., X-Ray Structure Determination: A PracticalGuide, Macmillan Co., New York (1968), Chapter 3, which is herebyincorporated by reference in its entirety. Alternatively, the uniquearrangement of atoms in spatial relation within the crystalline latticemay be characterized according to the observed fractional atomiccoordinates. Another means of characterizing the crystalline structureis by powder X-ray diffraction analysis in which the diffraction profileis compared to a simulated profile representing pure powder material,both run at the same analytical temperature, and measurements for thesubject form characterized as a series of 28 values.

One of ordinary skill in the art will appreciate that an X-raydiffraction pattern may be obtained with a measurement of error that isdependent upon the measurement conditions employed. In particular, it isgenerally known that intensities in an X-ray diffraction pattern mayfluctuate depending upon measurement conditions employed. It should befurther understood that relative intensities may also vary dependingupon experimental conditions, and, accordingly, the exact order ofintensity should not be taken into account. Additionally, a measurementerror of diffraction angle for a conventional X-ray diffraction patternis typically about 5 percent or less, and such degree of measurementerror should be taken into account as pertaining to the aforementioneddiffraction angles. Consequently, it is to be understood that thecrystal forms of the present disclosure are not limited to the crystalforms that provide X-ray diffraction patterns completely identical tothe X-ray diffraction patterns depicted in the accompanying Figuresdisclosed herein. Any crystal form that provides an X-ray diffractionpattern, and DSC thermogram substantially identical to those disclosedin the accompanying Figures fall within the scope of the presentdisclosure. The ability to ascertain substantial identities of X-raydiffraction patterns is within the purview of one of ordinary skill inthe art.

The present invention provides compositions containing thecompounds/crystalline forms described herein, including, in particular,pharmaceutical compositions comprising therapeutically effective amountsof the compounds/crystalline forms and pharmaceutically acceptablecarriers.

It is a further object of the present invention to provide kits having aplurality of active ingredients (with or without carrier) which,together, may be effectively utilized for carrying out the novelcombination therapies of the invention.

It is another object of the invention to provide a novel pharmaceuticalcomposition which is effective, in and of itself, for utilization in abeneficial combination therapy because it includes a plurality of activeingredients which may be utilized in accordance with the invention.

The present invention also provides kits or single packages combiningtwo or more active ingredients useful in treating the disease. A kit mayprovide (alone or in combination with a pharmaceutically acceptablediluent or carrier) the compounds of formula (I) or a crystalline formas described herein and the additional active ingredient (alone or incombination with diluent or carrier) selected from a serotonin 1Areceptor antagonist, a selective neurokinin-1 receptor antagonist, and anorepinephrine precursor.

In practice, the compounds/crystalline forms of the present inventionmay generally be administered parenterally, intravenously,subcutaneously, intramuscularly, colonically, nasally,intraperitoneally, rectally, or orally.

The products according to the present invention may be presented informs permitting administration by the most suitable route and theinvention also relates to pharmaceutical compositions containing atleast one product according to the invention which are suitable for usein human or veterinary medicine. These compositions may be preparedaccording to the customary methods, using one or more pharmaceuticallyacceptable adjuvants or excipients. The adjuvants comprise, inter alia,diluents, sterile aqueous media, and the various non-toxic organicsolvents. The compositions may be presented in the form of tablets,pills, granules, powders, aqueous solutions or suspensions, injectablesolutions, elixirs or syrups, and can contain one or more agents chosenfrom the group comprising sweeteners, flavorings, colorings, orstabilizers in order to obtain pharmaceutically acceptable preparations.

The choice of vehicle and the content of active substance in the vehicleare generally determined in accordance with the solubility and chemicalproperties of the product, the particular mode of administration and theprovisions to be observed in pharmaceutical practice. For example,excipients such as lactose, sodium citrate, calcium carbonate, dicalciumphosphate and disintegrating agents such as starch, alginic acids andcertain complex silicates combined with lubricants such as magnesiumstearate, sodium lauryl sulfate, and talc may be used for preparingtablets. To prepare a capsule, it is advantageous to use lactose andhigh molecular weight polyethylene glycols. When aqueous suspensions areused they can contain emulsifying agents or agents which facilitatesuspension. Diluents such as sucrose, ethanol, polyethylene glycol,propylene glycol, glycerol, and chloroform or mixtures thereof may alsobe used.

For parenteral administration, emulsions, suspensions, or solutions ofthe products according to the invention in vegetable oil, for examplesesame oil, groundnut oil, or olive oil, or aqueous-organic solutionssuch as water and propylene glycol, injectable organic esters such asethyl oleate, as well as sterile aqueous solutions of thepharmaceutically acceptable salts, are used. The solutions of the saltsof the products according to the invention are especially useful foradministration by intramuscular or subcutaneous injection. The aqueoussolutions, also comprising solutions of the salts in pure distilledwater, may be used for intravenous administration with the proviso thattheir pH is suitably adjusted, that they are judiciously buffered andrendered isotonic with a sufficient quantity of glucose or sodiumchloride, and that they are sterilized by heating, irradiation, ormicrofiltration.

Suitable compositions containing the compounds/crystalline forms of thepresent invention may be prepared by conventional means. For example,compounds/crystalline forms of the present invention may be dissolved orsuspended in a suitable carrier for use in a nebulizer or a suspensionor solution aerosol, or may be absorbed or adsorbed onto a suitablesolid carrier for use in a dry powder inhaler.

Solid compositions for rectal administration include suppositoriesformulated in accordance with known methods and containing at least onecompound of formula (I)/crystalline form.

The percentage of active ingredient in the compositions of the presentinvention may be varied, it being necessary that it should constitute aproportion such that a suitable dosage shall be obtained. Obviously,several unit dosage forms may be administered at about the same time.The dose employed will be determined by the physician, and depends uponthe desired therapeutic effect, the route of administration and theduration of the treatment, and the condition of the patient. In theadult, the doses are generally from about 0.01 to about 100 mg/kg bodyweight, preferably about 0.01 to about 10 mg/kg body weight per day byinhalation, from about 0.01 to about 100 mg/kg body weight, preferably0.1 to 70 mg/kg body weight, more especially 0.1 to 10 mg/kg body weightper day by oral administration, and from about 0.01 to about 50 mg/kgbody weight, preferably 0.01 to 10 mg/kg body weight per day byintravenous administration. In each particular case, the doses will bedetermined in accordance with the factors distinctive to the subject tobe treated, such as age, weight, general state of health, and othercharacteristics which can influence the efficacy of the medicinalproduct.

The products according to the present invention may be administered asfrequently as necessary in order to obtain the desired therapeuticeffect. Some patients may respond rapidly to a higher or lower dose andmay find much weaker maintenance doses adequate. For other patients, itmay be necessary to have long-term treatments at the rate of 1 to 4doses per day, in accordance with the physiological requirements of eachparticular patient. Generally, the active product may be administeredorally 1 to 4 times per day. It goes without saying that, for otherpatients, it will be necessary to prescribe not more than one or twodoses per day.

The present invention provides compounds which inhibit synapticnorepinephrine, dopamine, and serotonin uptake and are, therefore,believed to be useful in treating a disorder which is created by or isdependent upon decreased availability of serotonin, norepinephrine, ordopamine. Although the compounds of formula (I) inhibit synapticnorepinephrine, dopamine, and serotonin uptake, in any individualcompound, these inhibitory effects may be manifested at the same orvastly different concentrations or doses. As a result, the compounds offormula (I) are useful in treating such a disorder at doses at whichsynaptic norepinephrine uptake may be substantially inhibited but atwhich synaptic serotonin uptake or dopamine uptake is not substantiallyinhibited, or vice versa. Also, the compounds of formula (I) are usefulin treating such a disorder at doses at which synaptic dopamine uptakemay be substantially inhibited but at which synaptic norepinephrine orserotonin uptake is not substantially inhibited, or vice versa. And,conversely, the compounds of formula (I) are useful in treating such adisorder at doses at which synaptic serotonin uptake may besubstantially inhibited but at which synaptic norepinephrine or dopamineuptake is not substantially inhibited, or vice versa. The compounds offormula (I) may also be useful in treating such a disorder at doses atwhich synaptic norepinephrine, dopamine, and serotonin uptake aresubstantially inhibited.

The concentrations or doses at which a test compound inhibits synapticnorepinephrine, dopamine, and serotonin uptake is readily determined bythe use of standard assay and techniques well known and appreciated byone of ordinary skill in the art. For example, the degree of inhibitionat a particular dose in rats can be determined by the method of Dudley,J Pharmacol Exp Ther, 217:834-840 (1981), which is hereby incorporatedby reference in its entirety.

The therapeutically effective inhibitory dose is one that is effectivein substantially inhibiting synaptic norepinephrine uptake, synapticdopamine uptake, or synaptic serotonin uptake or inhibiting the synapticuptake of two or more of norepinephrine, dopamine, and serotonin uptake.The therapeutically effective inhibitory dose can be readily determinedby those skilled in the art by using conventional range findingtechniques and analogous results obtained in the test systems describedabove.

Compounds of this invention provide a particularly beneficialtherapeutic index relative to other compounds available for thetreatment of similar disorders. Without intending to be limited bytheory, it is believed that this is due, at least in part, to thecompounds having higher binding affinities for one or two of theneurotransmitter transporters, e.g., selectivity towards the serotonintransporter protein (“SERT”) over the transporters for otherneurochemicals, e.g., the dopamine transporter protein (“DAT”) and thenorepinephrine transporter protein (“NET”).

Binding affinities are demonstrated by a number of means well known toordinarily skilled artisans, including, without limitation, thosedescribed in the Examples section below. Briefly, for example,protein-containing extracts from cells, e.g., HEK293E cells, expressingthe transporter proteins are incubated with radiolabelled ligands forthe proteins. The binding of the radioligands to the proteins isreversible in the presence of other protein ligands, e.g., the compoundsof the present invention; said reversibility, as described below,provides a means of measuring the compounds' binding affinities for theproteins (IC₅₀ or Ki). A higher IC₅₀/Ki value for a compound isindicative that the compound has less binding affinity for a proteinthan is so for a compound with a lower IC₅₀/Ki; conversely, lowerIC₅₀/Ki values are indicative of greater binding affinities.

Accordingly, the difference in compound selectivity for proteins isindicated by a lower IC₅₀/Ki for the protein for which the compound ismore selective, and a higher IC₅₀/Ki for the protein for which thecompound is less selective. Thus, the higher the ratio in IC₅₀/Ki valuesof a compound for protein A over protein B, the greater is thecompounds' selectivity for the latter over the former (the former havinga higher IC₅₀/Ki and the latter a lower IC₅₀/Ki for that compound).

The compounds (“triple action transporter reuptake inhibitors”) of thepresent invention have potent binding affinity simultaneously for allthree of the biogenic amine transporters, NET, DAT, or SERT. Forexample, the compounds of this invention possess potent NET, DAT, & SERTIC₅₀/Ki values of less than 200 nM.

The in vivo affinity of the compounds to the three transporter proteins,SERT, DAT, and NET are demonstrated by means well known to those ofordinary skill in the art, including, without limitation, thosedescribed in the Examples section below.

Accordingly, the difference in compound selectivity in vivo for proteinis indicated by a higher percent occupancy value (or percent inhibitionof the [³H] ligand compound used in the Examples section) at thetransporter protein for which the compound is more selective, and alower percent occupancy (or percent inhibition of the ³[H] ligandcompound used in the Examples section) for the protein for which thecompound is less selective.

The compounds of the present invention, when administrated at apharmaceutically feasible dose via means such as, but not limited to,oral, intravenous, subcutaneous, intraperitoneal and intramuscular, havestatistically significant percent occupancy value(s) at one, two or allof the biogenic amine transporters NET, DAT, or SERT.

The compounds of the present invention, when administrated at apharmaceutically feasible dose via means such as, but not limited to,oral, intravenous, subcutaneous, intraperitoneal and intramuscular, have10%-100% occupancy value(s) at one, two or all of the biogenic aminetransporters NET, DAT, or SERT. In a preferred embodiment, compounds ofthe present invention have 40%-100% occupancy value(s) at at least onethe biogenic amine transporters NET, DAT, or SERT.

Examples Example 1 Preparation of7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline,L-Tartrate Salt

Step A: To a solution of 3-methoxybenzaldehyde (180 g, 1.32 mol) inmethanol (1 L) was added a 40% aqueous solution of methylamine (113 ml,1.31 mol) followed by 1 Hour stirring at 0° C. Sodium borohydride (75 g,1.98 mol) was added portionwise at 0° C. and the reaction mixture wasstirred for 1 Hour. The solution was concentrated to a smaller volumethen, was diluted with water (200 mL) and the resulting solution wasextracted with methylene chloride (3×500 mL). The combined organicextracts were dried over sodium sulfate, filtered, and concentratedunder reduced pressure to afford the crude N-methylbenzylamine (220 g,quantitative) as clear oil, which was used in the next step withoutfurther purification: ¹H NMR (CDCl₃, 500 MHz) δ.7.23 (t, J=8.0 Hz, 1H),6.92-6.88 (m, 2H), 6.81-6.78 (m, 1H), 3.80 (s, 3H), 3.73 (s, 3H), 2.07(broad s, 1H).

Step B: To a solution of the above amine (6.2 g, 41.00 mmol) from Step Ain methylene chloride (100 mL) was added 3,4-dichlorophenacyl bromide(10.0 g, 37.3 mmol) and the resulting mixture was stirred at 0° C. for 1Hour prior to the addition of triethylamine (5.20 mL, 37.31 mmol),followed by 1 Hour stirring at 0° C. The reaction mixture was dilutedwith water (100 mL) then the aqueous phase was extracted with additionalmethylene chloride (3×75 mL). The combined extracts were dried oversodium sulfate, filtered, and concentrated to afford1-(3,4-dichlorophenyl)-2-((3-methoxybenzyl)(methyl)amino)ethanone (15.08g) as a light yellow oil, which was used in the next step withoutfurther purification: ¹H NMR (500 MHz, CDCl₃) δ 8.08 (d, J=2.0 Hz, 1H),7.78 (dd, J=8.5; 2.0 Hz, 1H), 7.50 (d, J=8.5 Hz, 1H), 7.25 (d, J=8.5 Hz,1H), 6.90 (d, J=7.5 Hz, 1H), 6.87 (d, J=2.5 Hz, 1H), 6.82 (dd, J=8.0;2.5 Hz, 1H), 3.79 (s, 3H), 3.66 (s, 2H), 3.60 (s, 2H), 2.33 (s, 3H).

Step C: To a solution of the ketone (˜37 mmol) from Step B in methanol(150 mL), was added sodium borohydride (2.11 g, 55.79 mmol) portionwiseat 0° C. The reaction mixture was first stirred for 2 Hours then, wasdiluted with water (100 mL) and extracted with methylene chloride (3×300mL). The combined organic extracts were dried over sodium sulfate,filtered, and concentrated to dryness under reduced pressure to affordthe crude alcohol (14.14 g) as a yellow oil, which was used withoutfurther purification in the next step: ¹H NMR (500 MHz, CDCl₃) δ 7.45(d, J=2.0 Hz, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.28-7.23 (m, 1H), 7.16 (dd,J=8.0; 2.0 Hz, 1H), 6.90-6.81 (m, 3H), 4.70-4.65 (m, 1H), 3.81 (s, 3H),3.70 (d, J=13.0 Hz, 1H), 3.50 (d, J=13.0 H 1H), 2.54-2.49 (m, 2H), 2.32(s, 3H).

Step D: To a solution of the alcohol (˜37 mmol) from Step C in methylenechloride (200 mL) was added concentrated sulfuric acid (12 mL, 235 mol)and the mixture was stirred at 0° C. for 28 Hours. The reaction wasquenched by adding a 6N NaOH solution till pH˜9. The aqueous phase wasextracted with additional methylene chloride (3×). The combined organicextracts were washed with brine (3×), dried over sodium sulfate,filtered, and concentrated. The residue was purified by flashchromatography (1:1:1: to 1:1:2 dichloromethane/hexanes/ethyl acetate)to afford4-(3,4-dichlorophenyl)-7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline(7.0 g, 59% over 3 steps) as a light yellow oil: ¹H NMR (500 MHz, CDCl₃)δ 7.33 (d, J=8.0 Hz, 1H), 7.29 (d, J=2.0 Hz, 1H), 7.03 (dd, J=8.5; 2.0Hz, 1H), 6.76 (d, J=8.5 Hz, 1H), 6.66 (dd, J=8.5; 3.0 Hz, 1H), 6.61 (d,J=2.5 Hz, 1H), 4.16-4.11 (m, 1H), 3.77 (s, 3H), 3.67-3.59 (m, 2H), 2.92(dd, J=11.5; 5.5 Hz, 1H), 2.55 (dd, J=11.5; 7.0 Hz, 1H), 2.39 (s, 3H).The undesired 5-methoxy isomer was also isolated (1.20 g, 10% over 3steps).

Step E: The racemic4-(3,4-dichlorophenyl)-7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline(7.0 g) from Step D above was resolved by preparative chiral HPLC(CHIRALPAK AD column, using 80:20:0.1Heptane/2-propanol/diethylamine asthe eluent) to give the (+)-enantiomer ([α]²⁵ _(D)+31.9° (c 0.49,methanol)) (3.68 g) as a colorless oil and the (−)-enantiomer (3.99 g)as a colorless oil.

Step F: A solution of(+)-4-(3,4-dichlorophenyl)-7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline(3.68 g, 11.42 mmol) in a mixture of acetic acid (20 mL) and 48% aqueoushydrobromic acid solution (50 mL) was refluxed for 8 Hours. The ice-coldreaction mixture was basified with a concentrated aqueous solution ofsodium hydroxide and a saturated aqueous solution of sodium bicarbonateuntil reaching a pH of about 8-9 and was extracted with dichloromethane(3×). The combined extracts were dried over sodium sulfate, filtered,and concentrated in vacuo to afford the crude alcohol (2.6 g) as ayellow solid. ¹H NMR (500 MHz, CDCl₃) δ 7.32 (d, J=8.5 Hz, 1H), 7.26 (d,J=2.0 Hz, 1H), 7.01 (dd, J=8.0; 2.0 Hz, 1H), 6.65 (d, J=8.0 Hz, 1H),6.54 (d, J=8.5 Hz, 1H), 6.49 (broad s, 1H), 4.15-4.10 (m, 1H), 3.60 (d,J=15.0 Hz, 1H), 3.56 (d, J=15.0 Hz, 1H), 2.96 (dd, J=11.5; 5.7 Hz, 1H),2.52 (dd, J=11.5, 8.0 Hz, 1H), 2.39 (s, 3H).

Step G: To a solution of the phenol from Step F above (2.1 g, 6.81 mmol)and pyridine (0.72 mL, 8.85 mmol) in dichloromethane (60 mL) was addedtrifluoromethanesulfonic anhydride (1.37 mL, 8.14 mmol) at −78° C. Thereaction was allowed to warm to 0° C. and stirred for 1 Hour. Thereaction mixture was diluted with water (20 mL) and extracted withdichloromethane (3×). The combined extracts were dried over sodiumsulfate, filtered, and concentrated to give the crude triflate as ayellow oil. ¹H NMR (500 MHz, CDCl₃) δ 7.36 (d, J=8.5 Hz, 1H), 7.30 (d,J=2.0 Hz, 1H), 7.03-6.98 (m, 3H), 6.94 (d, J=8.5 Hz, 1H), 4.19-4.15 (m,1H), 3.68 (s, 2H), 2.96 (dd, J=11.7; 5.5 Hz, 1H), 2.60 (dd, J=11.7, 7.5Hz, 1H), 2.42 (s, 3H).

Step H: A mixture of the triflate from Step G above (˜6.8 mmol),bis(pinacolato)diboron (2.07 g, 8.15 mmol), and potassium acetate (2.05g, 20.8 mmol) in dimethyl sulfoxide (35 mL) was degassed with argon. Tothis mixture was addeddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (0.40 g,0.55 mmol). The resulting mixture was degassed with argon and thenheated at 85° C. for 2 Hours. The cold reaction mixture was diluted withethyl acetate (150 mL). The resulting solution was washed with water(2×40 mL), brine (1×40 mL), dried over sodium sulfate, filtered, andconcentrated in vacuo. A Purification flash chromatography column(eluent, 1:1:1 to 1:1:2 dichloromethane/hexanes/ethyl acetate) gave thedesired boronate ester (2.6 g, 91% over 2 steps) as a yellow solid. ¹HNMR (500 MHz, CDCl₃) δ 7.55 (s, 1H), 7.52 (d, J=7.5 Hz, 1H), 7.33 (d,J=8.5 Hz, 1H), 7.28 (d, J=2.0 Hz, 1H), 7.01 (dd, J=8.0, 2.0 Hz, 1H),6.85 (d, J=8.0 Hz, 1H), 4.23 (t, J=6.5 Hz, 1H), 3.71 (d, J=15.0 Hz, 1H),3.67 (d, J=15.0 Hz, 1H), 2.98 (dd, J=11.4, 5.3 Hz, 1H), 2.56 (dd,J=11.4, 7.5 Hz, 1H), 2.41 (s, 3H), 1.33 (s, 12H).

Step I: To a solution of the boronate ester (2.6 g, 6.22 mmol) from StepF and proton sponge (2.6 g, 12.1 mmol) in dichloroethane (80 mL) at 0°C. was added 1-chloroethyl chloroformate (2.4 mL, 22.1 mmol). Themixture was stirred at 0° C. for 15 minutes, then was refluxed for 40minutes and was concentrated in vacuo. The residue was filtered througha short pad of silica gel (eluent, 1:1:1 dichloromethane/hexanes/ethylacetate) and the filtrate was concentrated in vacuo. The residue wasdiluted with methanol (160 mL), heated to reflux for 1 Hour andconcentrated in vacuo to give the4-(3,4-dichlorophenyl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinolineas a brown foam.

Step J: A solution of the product from Step I (˜6.2 mmol), (Boc)₂O (3.60g, 16.4 mmol), triethylamine (1.5 mL, 10.7 mmol) and DMAP (0.26 g, 2.20mmol) in dichloromethane (120 mL) was stirred at room temperature for 4Hours. The reaction was quenched by the addition of water (50 mL) then,the aqueous phase was extracted with additional dichloromethane (2×100mL). The combined extracts were dried over sodium sulfate, filtered, andconcentrated in vacuo. A purification by flash column chromatography(eluent, 47.5:47.5:5 to 1:1:1 dichloromethane/hexanes/ethyl acetate)gave the boc-protected tetrahydroisoquinoline (1.82 g, 58% over 3 steps)as a white foam. ¹H NMR (500 MHz, CDCl₃) δ 7.65 (s, 1H), 7.58 (d, J=7.5Hz, 1H), 7.32 (d, J=8.0 Hz, 1H), 7.13 (s, 1H), 6.95 (d, J=7.0 Hz, 1H),6.97-6.93 and 6.83-6.78 (m, 1H), 5.01-4.95 and 4.48-4.43 (m, 1H),4.56-4.52 (m, 1H), 3.95 (s, 1H), 3.83-3.44 (m, 2H), 1.43 and 1.26 (2s,9H), 1.33 (s, 12H).

Step K: A dry flask was loaded with the boronate ester (0.8 g, 1.59mmol) from Step J, 6-bromo-[1,2,4]triazolo[1,5-a]pyridine (0.35 g, 1.78mmol), cesium carbonate (0.97 g, 2.98 mmol), anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)dichloromethane adduct (87 mg, 0.12 mmol). The flask was blanketed withargon then, DMF (20 mL) and water (4 mL) were added followed by a shortsonication. The reaction mixture was heated to 80° C. for 1 hour. Thecold reaction mixture was diluted with water (20 mL) and the aqueouslayer was extracted with dichloromethane (3×60 mL). The combined organicphases were concentrated in vacuo. Purification by flash columnchromatography (eluent, 1:1:1 to 1:1:2 dichloromethane/hexanes/ethylacetate) gave the Boc-protected7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline(0.86 g, quantitative) as a white foam.

Step L: A solution of the Boc-protected7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline(0.85 g, 1.72 mmol) and concentrated hydrochloric acid (4.0 mL) inethanol (10 mL) was stirred at room temperature for 1 Hour. The reactionmixture was concentrated to dryness in vacuo. The residue was dissolvedin a mixture of dichloromethane (14 mL) and TFA (10 mL), stirred at roomtemperature for 1 Hour then concentrated in vacuo. The syrup thusobtained was diluted with dichloromethane and treated with a saturatedaqueous solution of sodium bicarbonate until pH 8-9. The aqueous phasewas extracted with additional dichloromethane (3×) and the organicphases were dried over sodium sulfate, filtered, and concentrated invacuo togive7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline(0.59 g, 87%) as a white foam.

Step M: To a solution of the product (0.59 g, 1.49 mmol) from Step B inethanol was added L-tartaric acid (0.22 g, 1.49 mmol). The slurry wasfiltered. The cake was rinsed with ethanol and dried to give7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline,L-tartrate salt (0.49 g, 59%, AUC HPLC >99%) as a white solid. [[α]²⁵_(D)+9.0° (c 0.11, methanol)]. ¹H NMR (500 MHz, CD₃OD) δ 9.09 (s, 1H),8.53 (s, 1H), 8.02 (dd, J=9.0, 2.0 Hz, 1H), 7.86 (d, J=9.0 Hz, 1H), 7.68(s, 1H), 7.64-7.61 (m, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.48 (d, J=2.0 Hz,1H), 7.24 (dd, J=8.0, 2.0 Hz, 1H), 7.04 (d, J=8.0 Hz, 1H), 4.65-4.57 (m,2H), 4.52 (d, J=16.0 Hz, 1H), 4.41 (s, 2H), 3.79 (dd, J=12.5, 6.0 Hz,1H), 3.44 (t, J=12.5 Hz, 1H). ESI MS m/z 395 [M+H]⁺. Anal. Calcd. forC₂₁H₁₆Cl₂N₄.C₄H₆O₆.0.5H₂O: C, 54.16;H, 4.18; N, 10.11. Found: C, 54.07;H3.92; N, 9.97.

The L-tartrate of the(−)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinewas prepared using(−)-4-(3,4-dichlorophenyl)-7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolinefollowing similar steps described for the synthesis of the(+)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline,L-tartrate salt ([α]²⁴ _(D)−6.0° (c 0.10, methanol)).

Example 2 Alternate Synthesis of Example 1

Step A: To a solution of the triflate (9.5 g, 21.6 mmol) from step G inExample 1 and bis(pinacolato)diboron (6.6 g, 25.9 mmol) in dimethylsulfoxide (200 mL) was added potassium acetate (6.4 g, 64.8 mmol). Thesolution was degassed with argon for 5 minutes and thendichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (1.6 g, 2.2mmol) was added to it. The reaction mixture was degassed with argon for5 minutes, heated at 80° C. for 1 Hour, and then cooled to roomtemperature. To this solution were added6-bromo-[1,2,4]triazolo[1,5-α]pyridine (4.8 g, 23.8 mmol) and an aqueoussolution of cesium carbonate (21.1 g, 64.8 mmol in 87 mL of water). Theresultant solution was degassed with argon and thendichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) (0.8 g, 1.1mmol) was added to it. The reaction mixture was degassed with argon andheated at 80° C. for 1 Hour. A dark sticky oil formed during thereaction. The dark supernatant solution was poured out, diluted withwater, and extracted with ethyl acetate (3×), which was dried oversodium sulfate and concentrated in vacuo. The oil left was dissolved indichloromethane and the resultant solution was washed with water, driedover sodium sulfate, and concentrated in vacuo. The combined crudeproduct was purified by flash column chromatography (100% ethyl acetateto 92:7.2:0.8 ethyl acetate/methanol/ammonium hydroxide) to give7-([1,2,4]triazolo[1,5-α]pyridin-6-yl)-4-(3,4-dichlorophenyl)-2-methyl-1,2,3,4-tetrahydroisoquinoline(7.7 g, 87%, AUC HPLC 97.6%) as a brown foam: ¹H NMR (500 MHz, CDCl₃) δ8.77 (s, 1H), 8.37 (s, 1H), 7.82 (d, J=9.0 Hz, 1H), 7.76 (d, J=9.0 Hz,1H), 7.39-7.32 (m, 4H), 7.09 (d, J=8.0 Hz, 1H), 7.01 (d, J=8.5 Hz, 1H),4.26 (t, J=6.5 Hz, 1H), 3.75 (app s, 2H), 3.01 (dd, J=11.5, 5.5 Hz, 1H),2.64 (dd, J=11.5, 6.5 Hz, 1H), 2.46 (s, 3H).

Step B: To a solution of the7-([1,2,4]triazolo[1,5-α]pyridin-6-yl)-4-(3,4-dichlorophenyl)-2-methyl-1,2,3,4-tetrahydroisoquinoline(7.2 g, 17.6 mmol) from step A above in 1,2-dichloroethane (180 mL) at0° C. was added proton sponge (3.8 g, 17.6 mmol), followed by additionof 1-chloroethyl chloroformate (2.3 mL, 21.1 mmol). After the addition,the reaction solution was stirred at 0° C. for 20 minutes and roomtemperature for 14 Hours. Additional 1-chloroethyl chloroformate (0.5mL, 4.6 mmol) was added to the reaction solution. The reaction solutionwas stirred for another 3 Hours and then it was cooled to 0° C., washedwith aqueous hydrochloric acid (1N). Precipitate formed during the acidwash. The organic extract was separated, dried over sodium sulfate, andconcentrated in vacuo. The residue obtained was purified by flash columnchromatography (dichloromethane to 95:4.5:0.5dichloromethane/methanol/ammonium hydroxide) to give two batches ofpartially purified carbamate intermediates, which were dissolved inmethanol and refluxed for 1 Hour. The reaction solutions wereconcentrated in vacuo and the crude product obtained was purified by acombination of flash column chromatography (ethyl acetate to 88:10.2:0.8ethyl acetate/methanol/ammonium hydroxide) and preparative thin layerchromatography (ethyl acetate/methanol/ammonium hydroxide 90:9:1) togive the desired des-methyl tetrahydroisoquinoline (3.8 g, 54%; AUC HPLC98.7%) as a light pink foam: ¹H NMR (500 MHz, CDCl₃) δ 8.78-8.77 (m,1H), 8.37 (s, 1H), 7.83 (dd, J=9.5, 1.0 Hz, 1H), 7.77 (dd, J=9.0, 1.5Hz, 1H), 7.39 (d, J=8.5 Hz, 1H), 7.36-7.26 (m, 3H), 7.05-7.00 (m, 2H),4.24 (d, J=16.5 Hz, 1H), 4.17 (d, J=16.5 Hz, 1H), 4.13-4.11 (m, 1H),3.44 (dd, J=12.5, 5.0 Hz, 1H), 3.11 (dd, J=13.0, 6.0 Hz, 1H).

Step C: To a solution of des-methyl tetrahydroisoquinoline (3.75 g, 9.48mmol) from step B above in ethanol (80 mL) was added activated carbon(3.0 g) and stirred at room temperature for 30 minutes. The carbon wasremoved by filtration and the filtrate obtained was concentrated invacuo. The resultant oil was dissolved in ethanol (60 mL) and a solutionof L-tartaric acid (1.44 g, 9.5 mmol) in ethanol (20 mL) was added. Uponwhich, white precipitate formed immediately. The slurry was stirred atroom temperature for 10 minutes and filtered. The cake obtained wasstirred in hot ethanol (70° C.) for 3 Hours and filtered. The cakeobtained was dried in vacuo at 50-60° C. for 40 Hours to give the(+)-7-([1,2,4]triazolo[1,5-α]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolineL-tartrate (3.7 g, 73%; AUC HPLC 99.4% at 250 nm) as an off-white solid[α]²³ _(D)+16.8° (c 0.13, methanol): ¹H NMR (500 MHz, CD₃OD) δ 9.09 (s,1H), 8.53 (s, 1H), 8.02 (dd, J=9.0; 2.0 Hz, 1H), 7.86 (d, J=9.0 Hz, 1H),7.68 (s, 1H), 7.64-7.61 (m, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.48 (d, J=2.0Hz, 1H), 7.24 (dd, J=8.0; 2.0 Hz, 1H), 7.04 (d, J=8.0 Hz, 1H), 4.65-4.57(m, 2H), 4.52 (d, J=16.0 Hz, 1H), 4.41 (s, 2H), 3.79 (dd, J=12.5; 6.0Hz, 1H), 3.44 (t, J=12.5 Hz. 1H). ESI MS m/z 395 [M+H]⁺ Anal. Calcd. forC₂₁H₁₆Cl₂N₄.C₄H₆O₆.0.5H₂O; C, 54.16; H, 4.18; N, 10.11. Found: C,53.96;H 3.98; N, 9.94.

Example 3 Alternative Synthesis of Example 1 (Hydrochloride)

Step A: To a 1 L round-bottom flask was added 2-amino-5-bromopyridine(100 g, 578 mmol), DMF-DMA (101 mL, 751 mmol) and 2-propanol (200 mL).The mixture was heated to reflux for 3H to give a clear dark solution.It was then cooled to 50° C. and hydroxylamine hydrochloride (52.2 g,751 mmol) was added. The mixture was stirred at 50° C. overnight to givea yellow suspension. The precipitate was collected by filtration. Theblack filtrate was concentrated and the residue was stirred in EtOH (20mL) for 20 min. The solid was collected by filtration. The combinedsolids were dried in an oven to giveN-(5-bromopyridin-2-yl)-N′-hydroxyformimidamide as a sandy solid (94 g,75% yield).

Step B: N-(5-bromopyridin-2-yl)-N′-hydroxyformimidamide was dissolved inTHF (1 L). To the solution at 10° C. was added trifluoroacetic anhydride(106 mL, 751 mmol) slowly to control the reaction temperature below 20°C. After the addition was complete, the mixture was warmed to roomtemperature and stirred for 2H. After the reaction was finished, it wasquenched with Na₂CO₃ aqueous solution to adjust pH>7. The organicsolvent was removed under reduced pressure, and the product was thenextracted with DCM (4×300 mL). The combined organic layers were driedover Na₂SO₄ and concentrated to dryness. The residue was stirred inethyl ether (100 mL) and the product6-bromo-[1,2,4]triazolo[1,5-a]pyridine was collected by filtration as anoff-white solid (50 g, 58% yield).

Step C: To a mixture of 3-formylphenylboronic acid (21.41 g, 143 mmol),6-bromo-[1,2,4]triazolo[1,5-a]pyridine (28.27 g, 143 mmol) in DMSO (600mL) and water (50 mL) was added Pd(dppf)Cl₂ (5.83 g, 7.14 mmol) andCs₂CO₃ (116 g, 357 mmol). The reaction temperature reached 45° C. afterthe addition. HPLC showed that starting materials were consumed after 15min. The reaction was diluted with water (400 mL). The black precipitatewas collected by filtration and dissolved in DCM (300 mL), and washedwith brine (200 mL). The aqueous layer was back extracted with DCM (100mL). The combined organic layers were filtered through a Celite pad andthe filtrate was concentrated to give a black solid mixture. The productwas recrystallized in methanol to give3-([1,2,4]triazolo[1,5-a]pyridin-6-yl)benzaldehyde (27.4 g, 123 mmol,86% yield) as a pale grey solid: m/z=224.0 [M+1]; ¹H NMR (400 MHz,DMSO-D6) δ ppm 7.74 (t, J=7.68 Hz, 1H), 7.91-8.02 (m, 2H), 8.11 (dd,J=9.19, 1.89 Hz, 1H), 8.17 (d, J=7.81 Hz, 1H), 8.36 (s, 1H), 8.57 (s,1H), 9.45 (s, 1H), 10.11 (s, 1H).

Step D: A mixture of α-bromo-3,4′-dichloroacetophenone (26.7 g, 100mmol), hexamethylenetetramine (HMTA) (13.97 g, 100 mmol) and NaI (0.5 g)was stirred at room temperature overnight. HPLC analysis indicatedconsumption of starting materials. The ammonium intermediate wascollected by filtration as a white solid, washed with acetone and dried(36 g, 89% yield).

To a solution of the intermediate (36 g, 88 mmol) in EtOH (500 mL) wasadded 12 N HCl (75 mL, 0.9 mol). The mixture was stirred at 76° C.overnight, and then cooled to room temperature. The product2-amino-1-(3,4-dichlorophenyl)ethanone hydrochloride was obtained as acrystal solid by filtration (20.2 g, 95% yield): ¹H NMR (400 MHz,DMSO-D6) δ ppm 4.62 (s, 2H), 7.79-7.94 (m, 1H), 7.98 (dd, J=8.56, 2.01Hz, 1H), 8.26 (d, J=2.01 Hz, 1H), 8.48 (s, 3H).

Step E: To a solution of 2-amino-1-(3,4-dichlorophenyl)ethanonehydrochloride (50 g, 208 mmol) in MeOH (200 mL) was added sodiumborohydride (7.86 g, 208 mmol) at 0° C. slowly. HPLC indicated 100%conversion after 10 min. A solution of3-([1,2,4]triazolo[1,5-a]pyridin-6-yl)benzaldehyde (46.4 g, 208 mmol) inDCM/MeOH (180 mL/50 mL) was added to the previous solution in oneportion at room temperature. The mixed solution was stirred at RT for 2H, then sodium borohydride (7.86 g, 208 mmol) was added. HPLC indicated100% conversion after 10 min. Most of the solvent was removed and theresidual was dissolved in DCM/NH₄OH (4N) (1 L/1 L). The organic layerwas washed with brine, dried over Na₂SO₄, and concentrated to ˜250 mL.The product2-(3-([1,2,4]triazolo[1,5-a]pyridin-6-yl)benzylamino)-1-(3,4-dichlorophenyl)ethanolin DCM solution was used in the next step without further purification(HPLC area 92%): m/z=413.1 [M+1]; ¹H NMR (400 MHz, CHLOROFORM-D) δ ppm2.72 (dd, J=12.21, 8.69 Hz, 1H), 2.96 (dd, J=12.34, 3.53 Hz, 1 H ),3.85-3.98 (m, 2H), 4.69 (dd, J=8.56, 3.53 Hz, 1H), 7.18 (dd, J=8.31,1.76 Hz, 1 H), 7.34-7.42 (m, 2H), 7.43-7.56 (m, 4H), 7.72-7.88 (m, 2H),8.36 (s, 1H), 8.78 (s, 1H).

Step F: A solution of concentrated sulfuric acid (500 g, 5.0 mol) in a 3L round bottom flask was cooled to 0° C. with an ice bath. To the flaskwas added dropwise a solution of2-(3-([1,2,4]triazolo[1,5-a]pyridin-6-yl)benzylamino)-1-(3,4-dichlorophenyl)ethanol(79 g, 0.191 mol) in DCM (250 mL). The addition was finished in 30 minand the reaction temperature was controlled in the range of 10-20° C.DCM was blown away with nitrogen gas during the addition. Theevaporation of DCM helped to lower the reaction temperature. The mixturesolution was stirred at RT overnight. HPLC indicated no remainingstarting material. The HPLC area ratio of7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolineand5-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinewas 75:25. The reaction mixture was cooled to 0° C. Isopropanol (2 L)was added to the solution slowly, maintaining temperature <0° C. Thesolid (desired isomer 92% purity) was obtained by filtration. The solidwas then dissolved in AcOEt (1 L) and the pH adjusted to 10 with NH₄OH.The water layer was extracted with EtOAc twice. The combined organiclayers were washed with water, dried over Na₂SO₄ and concentrated. Theresidue was dissolved in EtOH (250 mL) and then 1.1 eq ofmethanesulfonic acid (20.20 g, 0.21 mol) was added and the solutionstirred overnight. The resulting precipitate methanesulfonic acid salt(98% purity) was filtered. This was dissolved in water and the pHadjusted with NH₄OH to 10, then extracted with AcOEt twice. The combinedextracts were washed with water and dried over Na₂SO₄. After removal ofsolvent,7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinewas obtained in an amorphous state (40.8 g, 54% yield): m/z=395.0 [M+1];¹H NMR (400 MHz, CHLOROFORM-D) δ ppm 3.05 (dd, J=12.00, 8.00 Hz, 1H),3.40 (dd, J=12.00, 4.00 Hz, 1H), 4.05-4.25 (m, 3H), 6.96 (m, 2H),7.25-7.35 (m, 4H), 7.70-7.80 (m, 2H), 8.32 (s, 1H), 8.74(s, 1H).

Step G: To a solution of7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline(25.2 g, 63.8 mmol) in DMF (30 ml) was added di-tert-butyl dicarbonate(13.91 g, 63.8 mmol). The reaction mixture was stirred at RT for 1 H,then AcOEt (500 ml) was added. The solution was washed with brine andwater. The organic layer was dried over Na₂SO₄. After removal ofsolvent, solid rac-tert-butyl7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate(30.6 g, 61.8 mmol, 97% yield) was obtained by recrystallization fromMeOH; m/z=495.1 [M+1]; ¹H NMR (400 MHz, CHLOROFORM-D) δ ppm 1.30 (s,9H), 3.60-4.15 (m, 3H), 4.40-5.10 (m, 2H), 6.84-7.05 (m, 2H), 7.13 (d,J=1.51 Hz, 1H), 7.35 (m, 3H), 7.78 (dd, J=8.31, 1.77 Hz, 2H), 8.31 (s,1H), 8.72 (s, 1H).

Step H: Chiral SFC separation on a Chiralpak AS-H column (3×25 cm, 5 μm;eluent: CO2/(MeOH/TEA=100/0.2(v/v))=75/25; 220 nm) yielded(+)-tert-butyl7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate(99.7% ee).

Step I: To a solution of the (+)-enantiomer from Step H (32.41 g, 65.43mmol) in DCM (150 ml) was added hydrogen chloride-EtOH solution (2.5N,250 mL) and EtOH 500 mL. The reaction mixture was stirred at 70° C. for2 h. After removal of the solvent, the residue was refluxed in 1000 mlAcOEt for 1 h. The product(+)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinehydrochloride (27.4 g, 97% yield) was obtained after filtration anddrying. m/z=395.1 [M+1]; ¹H NMR (400 MHz, DMSO-d6) δ ppm 3.70 (m, 2H),4.40-4.65 (m, 3H), 6.90 (d, 7.80 Hz, 1H), 7.35 (dd, J =7.8, 2 Hz, 1H),7.68 (m, 4H), 8.58 (s, 1H), 9.38 (s, 1H), 9.8 (bs, 2H).

Example 4 Primary Binding Assay Preparation of Membranes

Recombinant HEK-293 cells expressing either the hSERT, hDAT, or hNETproteins were harvested from T-175 flasks as follows. The medium wasremoved from the flasks and the cells rinsed with HBSS without Ca andwithout Mg. The cells were then incubated for 5-10 minutes in 10 mMTris-Cl, pH 7.5, 5 mM EDTA before the cells were lifted with acombination of pipetting and scraping, as needed. The cell suspensionwas collected into centrifuge bottles and homogenized for 30 secondswith a Polytron homogenizer. The suspension was centrifuged for 30minutes at 32,000×g, 4° C. The supernatant was decanted and the pelletresuspended and homogenized in 50 mM Tris-Cl, pH 7.5, 1 mM EDTA for 10seconds. The suspension was then centrifuged again for 30 minutes at32,000×g, 4° C. The supernatant was decanted and the pellet resuspendedin 50 mM Tris-Cl, pH 7.5, 1 mM EDTA and briefly homogenized. A Bradfordassay (Bio-rad) was performed and the membrane preparation diluted to 2mg/ml with 50 mM Tris-Cl, pH 7.5, 1 mM EDTA. Aliquots were prepared, andthen frozen and stored at −80° C.

SERT Radioligand Binding Assay

Compounds were dissolved in 100% DMSO at a concentration 100 times thedesired highest assay concentration, serially diluted 1:3 in 100% DMSO,and 0.4 μl/well of each solution was dispensed to a Nunc polypropylene,round bottom, 384-well plate. 100% inhibition is defined with 0.4μl/well of 1 mM fluoxetine dissolved in DMSO. 20 μl/well of a 2×membrane preparation (15 ug/ml in 50 mM Tris-Cl, pH 7.5, 120 mM NaCl, 5mM KCl) and 20 μl/well of a 2× radioligand solution (520 pM [¹²⁵I]RTI-55in 50 mM Tris-Cl, pH 7.5, 120 mM NaCl, 5 mM KCl) were added to each welland the reaction incubated for 1 Hour at room temperature. The contentsof the assay plate were then transferred to a MilliporeMultiscreen_(HTS) GF/B filter plate which was pretreated with 0.5% PEIfor at least one hour. The plate was vacuum filtered and washed with 7washes of 100 μl/well 50 mM Tris-Cl, pH 7.5, 120 mM NaCl, 5 mM KClchilled to 4° C. The filtration and washing were completed in less than90 seconds. The plates were air-dried overnight, 12 μl/well ofMicroScint scintillation fluid added, and the plates counted in aTrilux.

DAT Radioligand Binding Assay

Compounds were dissolved in 100% DMSO at a concentration 100 times thedesired highest assay concentration, serially diluted 1:3 in 100% DMSO,and 0.4 μl/well of each solution was dispensed to a Nunc polypropylene,round bottom, 384-well plate. 100% inhibition is defined with 0.4μl/well of 1 mM GBR-12935 dissolved in DMSO. 20 ul/well of a 2× membranepreparation (12.5 μg/ml in 30 mM sodium phosphate buffer, pH 7.9 at 4°C.) and 20 μl/well of a 2× radioligand solution (250 pM [¹²⁵I]RTI-55 in30 mM sodium phosphate buffer, pH 7.9 at 4° C.) were added to the welland the reaction incubated for 1 Hour at room temperature. The contentsof the assay plate were then transferred to a MilliporeMultiscreen_(HTS) GF/B filter plate which was pretreated with 0.5% PEIfor at least one hour. The plate was vacuum-filtered and washed with 7washes of 100 μl/well 50 mM Tris-Cl, pH 7.5, 120 mM NaCl, 5 mM KClchilled to 4° C. The filtration and washing were completed in less than90 seconds. The plates were air-dried overnight, 12 μl/well ofMicroScint scintillation fluid added, and the plates counted in aTrilux.

NET Radioligand Binding Assay

Compounds were dissolved in 100% DMSO at a concentration 100 times thedesired highest assay concentration, serially diluted 1:3 in 100% DMSO,and 1.0 μl/well of each solution was dispensed to a Nunc polypropylene,round bottom, 384-well plate. 100% inhibition is defined with 1.0μl/well of 10 mM desipramine dissolved in DMSO. 50 μl/well of a 2×membrane preparation (0.4 mg/ml in 50 mM Tris-Cl, pH 7.5, 120 mM NaCl, 5mM KCl) and 50 μl/well of a 2× radioligand solution (4 nM [³H]nisoxetinein 50 mM Tris-Cl, pH 7.5, 120 mM NaCl, 5 mM KCl) were added to the welland the reaction incubated for 1 Hour at room temperature. The contentsof the assay plate were then transferred to a MilliporeMultiscreen_(HTS) GF/B filter plate which was pretreated with 0.5% PEIfor at least one hour. The plate was vacuum filtered and washed with 7washes of 100 μl/well 50 mM Tris-Cl, pH 7.5, 120 mM NaCl, 5 mM KClchilled to 4° C. The filtration and washing were completed in less than90 seconds. The plates were air-dried overnight, 12 μl/well ofMicroScint scintillation fluid added, and the plates counted in aTrilux.

Data Analysis

The raw data was normalized to percent inhibition using control wellsdefining 0% (DMSO only) and 100% (selective inhibitor) inhibition whichwere run on each plate. Each plate was run in triplicate, and theconcentration response curve thus generated was fit using thefour-parameter dose response equation, Y=Bottom+(Top−Bottom)/(1+10̂((LogIC₅₀−X)*HillSlope)) in order to determine the IC₅₀ value for eachcompound. The radioligand concentration chosen for each assaycorresponds to the K_(d) concentration determined through saturationbinding analysis for each assay.

Example 5 Occupancy Assay

The general procedure for brain tissue collection and transporteroccupancy assessment is briefly described as follows. Mice weresacrificed by asphyxiation in CO₂, rats by decapitation and dogs by IVinjection of euthanasia solution. For mice and rats, after the brainswere removed from the skull, the forebrain tissue (removal of thebrainstem and cerebellum) was used for SERT, NET, and DAT occupancyassessment. In dogs, the striatum was dissected for DAT occupancy andthe remaining forebrain tissue (without the striatum, brainstem, andcerebellum) was used for SERT and NET occupancy assessment. The braintissues were frozen in chilled isopentane and stored at −80° C. untilhomogenization.

The brain tissues were thawed and then homogenized using a polytronhomogenizer (Kinematica). Sample aliquots were frozen immediately andstored at −80° C. Protein content was measured for each sample using aCoomassie protein assay kit (Pierce).

On the day of ex vivo binding for occupancy assessment, frozen samplealiquots were thawed and needle homogenized, and 100 μg of the tissuewas incubated for SERT, NET, and DAT binding under assay conditionssummarized in Table 1. After incubation, the reactions were terminatedby the addition of ice-cold assay buffer and rapid filtration through aBrandel Cell Harvester using FPXLR-196 filters. The filters were washedtwice with ice-cold incubation buffer, punched into a clear plate priorto the addition of 200 ul scintillation fluid per well. Radioligand wasmeasured using a Wallac Microbeta liquid scintillation counter.

TABLE 1 Ex Vivo Binding Assay Conditions for Serotonin, Norepinephrineand Dopamine Transporter Occupancy. Non- Incubation Specific Buffer Timeand Transporter Radioligand Drug (μM) (nM) Temperature SERT 2 nMFluoxetine, Tris, 50 10 minutes [³H]Citalopram 10 NaCl, 120 at 4° C.KCl, 5 DAT 0.1 nM GBR-12935, Sodium 10 minutes [¹²⁵I]RTI-55 10 phosphateat 4° C. (+0.5 μM buffer, 30 citalopram) NET 5 nM Reboxetine, Tris, 5020 minutes [³H]-Nisoxetine 10 NaCl, 300 at 4° C. KCl, 5

The specific binding was calculated by subtracting the value of thenon-specific binding from that of the total binding in each sample. Thepercent occupancy was calculated as (1-specific binding in drugtreated/specific binding in vehicle treated)×100%. For estimation of invivo occupancy EC₅₀ (total plasma concentration of compound producing50% occupancy), plots of occupancy values versus plasma concentrationswere fitted to a one-site binding model using nonlinear regressionaccording to the following equation: % Occupancy=Emax*C/(EC₅₀+C) whereEmax is the maximal specific binding, C is the drug concentration, andEC₅₀ is the total plasma concentration required for 50% binding siteoccupancy. Nonlinear regression was performed using GraphPad Prismversion 3.00 (GraphPad Software, San Diego, Calif.).

The results are shown in Table 2, below:

TABLE 2 IC₅₀ and Occupancy Values time SERT DAT NET SERT DAT NET pointExample IC50 (nM) IC50 (nM) IC50 (nM) Occupancy % Occupancy % Occupancy% Dose_mg/kg (hour) 1 (+)-enantiomer 1.8 30.8 26.0 75 26 11 1 3 1(−)-enantiomer 4.2 104.6 4.8 41 30 33 1 3

Example 6 In Vivo Behavioral Assays For All Tests

All animals were maintained in accordance with the guidelines of theCommittee on Animals of the Bristol-Myers Squibb Company and Guide forCare and Use of Laboratory Animals, Institute of Animal LaboratoryResources, 1996, which are hereby incorporated by reference in theirentirety. Research protocols were approved by the Bristol-Myers SquibbCompany Institutional Animal Care and Use Committee.

Mouse Tail Suspension Assay

Male Swiss Webster mice were housed 3-4 per cage in rooms maintained ata constant temperature (21-23° C.) and humidity (50±10%) on a 12-hourlight/dark cycle. Animals had ad libitum access to water and foodthroughout studies. On the day of testing, they were brought into thetesting room and allowed to acclimate for 1 Hour. To begin testing, thetail was attached to a piece of tape which was then attached to a hookon the ceiling of a sound-attenuated chamber. Immobility wasautomatically recorded using the Med Associates software. Compounds wereadministered acutely at a fixed pretreatment interval before session.

The minimum effective dose of Example 1-(+)-enantiomer in the mouse tailsuspension study was 10 mg/kg.

Rat Forced Swim Assay

Male Sprague Dawley rats are housed in pairs in rooms maintained at aconstant temperature (21-23° C.) and humidity (50±10%) on a 12-hourlight/dark cycle. Animals have ad libitum access to water and foodthroughout studies. Animals are handled for two minutes each on the twodays prior to the start of the experiment. On the first day of testing,rats are placed in the swim tank (a Pyrex cylinder 46 cm tall×21 cm indiameter, filled with 30 cm of water ranging between 24-26° C.) for 15minutes (the pre-swim session). At the end of the 15-minute session,rats are dried and replaced in their home cage. Compounds areadministered at three time points in the next 24 Hour (23.5, 5, and 1Hour), prior to a second test swim. This swim test is 5 minutes induration and the animals' behavior is videotaped and active behaviors(immobility, swimming, climbing) are scored. At the end of each 5-secondperiod during the 5-minute test session the rat's behavior is scored asone of the following: immobility (the rat remained floating in the waterwithout struggling and made only those movements necessary to keep itshead above water), swimming (the rat made active swimming motions, morethan necessary to merely maintain its head above water, e.g., movingaround in the cylinder), or climbing (the rat made active movements withits forepaws in and out of the water, usually directed against thecylinder wall). Compounds are only identified by a predesignated codeand the experimenter remains blinded throughout the experiment(including while scoring videotapes).

Rat and Mouse Locomotor Activity

Animals are housed according to conditions described above for the twospecies. The testing apparatus consists of Plexiglas chambers equippedwith Digiscan activity monitors (Omnitech Electronics, Columbus, Ohio)that detect interruptions of eight photobeams. Horizontal activity isrecorded in 5-minute bins for a total of 60 minutes and expressed astotal distance covered (in cm). Compounds are administered acutely at afixed pretreatment interval prior to testing.

Example 7 Preparation of single crystals of(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolineL-tartrate (L-tartrate salt)

(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolineL-tartrate salt (20 mg) was dissolved in methanol (8 mL) under heatingin a vial. Distilled water (2 mL) was then added to the above clearsolution. The resulting solution was capped and placed at roomtemperature. Needle-like crystals of(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolineL-tartrate salt were obtained after slow evaporation in air within days.

Example 8 Preparation of single crystals of(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemonohydrochloride monoisopropanolate monohydrate (HCl salt; Form SA-1)

(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemono-HCl salt (20 mg) was dissolved in isopropanol (10 mL) under heatingin a vial. Distilled water (2 mL) was then added to the above clearsolution. The resulting solution was capped and placed at roomtemperature. Long needle crystals of(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemono-HCl monoisopropanolate monohydrate salt were obtained after slowevaporation in air within days. The needle-like crystals were separatedfrom mother liquor by filtration and the wet cake was dried in an ovenfor 16 hours under the condition of 45° C. and 100 mmHg.

Example 9 Preparation of single crystals of(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemonohydrochloride (HCl salt; Form N-2)

(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemono-HCl salt (20 mg) was dissolved in methanol (8 mL) under heating ina vial. Distilled water (2 mL) was then added to the above clearsolution. The resulting solution was capped and placed at roomtemperature. Needle like single crystals of(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemono-HCl salt were obtained after slow evaporation in air within days.

Example 10 Single Crystal Analysis by X-Ray Crystallography

The data of(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolineL-tartrate (L-tartrate salt) and(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemonohydrochloride (HCl salt; Form N-2) crystals were collected on aSMART CCD diffractometer equipped with graphite-monochromated Cu Kαradiation (λ=1.54178 Å) at 225K and the room temperature, respectively.The data of(S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemonohydrochloride monoisopropanolate monohydrate (HCl salt; Form SA-1)were collected on an X8-ApexII diffractometer equipped withgraphite-monochromated Cu Kα radiation (λ=1.54178 Å) at room temperature(APEX-II 1.0-28, Data Collection Software for Bruker CCD devices. BrukerAXS Inc., Madison, Wis., US. SAINT PLUS, Processing Software forBrukerCCD devices, Bruker AXS Inc., Madison, Wis., US). The final unitcell parameters were determined using the entire data set.

All structures were solved by direct methods and refined by thefull-matrix least-squares techniques, using the SHELXTL software package(Sheldrick, GM. 1997, SHELXTL. Structure Determination Programs. Version5.10, Bruker AXS, Madison, Wis., USA.). The function minimized in therefinements was Σ_(w)(|F_(o)|−|F_(c)|)². R is defined asΣ∥F_(o)|−|F_(c)∥/Σ|F_(o)| whileR_(w)=[Σ_(w)(|F_(o)|−|F_(c)|)2/Σ_(w)|F_(o)|²]^(1/2), where w is anappropriate weighting function based on errors in the observedintensities. Difference Fourier maps were examined at all stages ofrefinement. In L-tartrate form, one of chloro atoms on pendant phenylring is disordered over two positions with 50% occupancy ratio each. Thetartaric acid molecule is also disordered, which could not be modeledwell. The numbers of methanol molecules could not be identified due todisorder. All non-hydrogen atoms were refined with anisotropic thermaldisplacement parameters. The hydrogen atoms associated with hydrogenbonding were located in the final difference Fourier maps while thepositions of the other hydrogen atoms were calculated from an idealizedgeometry with standard bond lengths and angles. They were assignedisotropic temperature factors and included in structure factorcalculations with fixed parameters.

The crystal data of the L-tartrate salt form is shown in Table 3 and thefractional atomic coordinates are listed in Table 4. The crystal data ofForm SA-1 is shown in Table 5 and the fractional atomic coordinates arelisted in Table 6. The crystal data of Form N-2 is shown in Table 7 andthe fractional atomic coordinates are listed in Table 8. It should beunderstood by one of ordinary skills in the art that slight variationsin the coordinates are possible and are considered to be within thescope the present disclosure.

TABLE 3 Crystal Data of L-tartrate Form Empirical formula C40 H40 Cl2 N8O8 Formula weight 831.70 Temperature 225(1) K Wavelength 1.54178 ÅCrystal system, space group Orthorhombic, C222₁ Unit cell dimensions a =7.6264(10) Å alpha = 90 deg. b = 38.942(5) Å beta = 90 deg. c =24.449(3) Å gamma = 90 deg. Volume 7261.1(16) Å³ Z, Calculated density8, 1.522 Mg/m³ Absorption coefficient 2.195 mm⁻¹ F(000) 3472 Theta rangefor data collection 2.27 to 66.20 deg. Limiting indices −8 <= h <= 8,−45 <= k <= 42, −22 <= l <= 28 Reflections collected/unique 24815/6156[R(int) = 0.1027] Refinement method Full-matrix least-squares onF{circumflex over ( )}2 Data/restraints/parameters 6156/2/323Goodness-of-fit on F{circumflex over ( )}2 2.340 Final R indices [I > R1= 0.2345, wR2 = 0.4418 2sigma(I)] R indices (all data) R1 = 0.3127, wR2= 0.4595 Absolute structure parameter 0.00(11) Extinction coefficient0.0075(9) Largest diff. peak and hole 0.991 and −0.773 e. Å⁻³

TABLE 4 Atomic Coordinates of L-tartrate Form Atomic coordinates (×10⁴)and equivalent isotropic displacement parameters (Å² × 10³) forL-tartrate Form. x y z U(eq) Cl(1) 8174(9)    94(1) 4057(2) 171(2)Cl(2′) 5256(12)  −323(2) 4561(3) 137(3) Cl(2) 11696(16)   −83(2) 4303(4)201(6) C(1) 8480(30)  −296(3) 4374(6) 109(5) C(2) 10110(40)   −377(4)4452(8) 149(8) C(3) 10610(20)   −698(5) 4682(7) 136(6) C(4) 9280(20) −919(2) 4902(4)  78(3) C(5) 7540(20)  −803(3) 4839(5) 107(4) C(6)7210(20)  −477(3) 4556(5) 109(5) C(7) 9651(19) −1252(2) 5194(5)  97(4)C(8) 8790(20) −1532(3) 4886(5) 122(5) C(9) 7840(20) −1835(2) 5751(6)111(5) C(10) 8275(16) −1504(3) 6055(6)  87(3) C(11) 9041(16) −1238(2)5781(5)  83(3) C(12) 9409(14)  −941(2) 6125(5)  71(3) C(13) 8887(15) −937(3) 6658(6)  82(3) C(14) 8050(16) −1194(3) 6915(5)  75(3) C(15)7808(18) −1500(2) 6586(6)  90(4) C(16) 7563(15) −1182(2) 7472(6)  79(3)C(17) 6993(17)  −875(4) 7699(6)  96(4) C(18) 6487(18) −1113(4) 8577(8)100(4) C(19) 7058(19) −1442(5) 8390(5) 112(5) C(20) 7492(19) −1472(3)7861(7) 118(5) C(21) 5610(30)  −748(9) 8994(6)  194(13) C(22) 7820(20)−2663(4) 4481(6) 124(4) O(3) 10030(30)  −2275(4) 4338(6) 225(7) C(23)9000(20) −2557(4) 4090(6) 119(4) O(2) 7170(20) −2487(3) 4903(5) 170(4)O(1) 7230(20) −2972(3) 4484(5) 186(5) N(1) 8830(20) −1870(2) 5245(6)138(5) N(2) 6491(14)  −849(3) 8247(6) 109(4) N(3) 5890(20) −1046(4)9099(9) 150(7) N(4) 5882(18)  −566(3) 8552(6) 119(4) O(8) −840(20)  53(4) 2431(8) 235(7) O(1W) 9327(17) −3528(3) 4909(5) 175(4) C(74) 450(50) −1233(9)  3340(13)  272(14) O(9) −2350(140)  −964(16)  3320(30) 630(40) O(4) 7600(60) −2153(9)  3690(14)  400(15) O(6) 10620(40) −2645(6) 3106(9) 291(9) C(72) −2920(80)   −1321(14)  3380(20)  400(30)O(7) −160(50)  −761(8)  3131(12)  351(13) C(70)  −300(120)  −361(12) 2710(20)  420(30) C(25) 9840(80)  −2305(16)  3320(20)  440(30) O(5)8080(40) −2558(7) 2969(9)  312(11) C(24) 8360(40) −2552(8)  3522(10) 241(11) H(3A) 11778 −764 4690 164 H(5A) 6612 −931 4976 128 H(7A) 10920−1291 5191 116 H(8A) 9408 −1570 4544 146 H(8B) 7592 −1469 4803 146 H(9A)8097 −2030 5986 133 H(9B) 6598 −1839 5669 133 H(12A) 10003 −753 5980 85H(13A) 9130 −740 6861 99 H(15A) 7325 −1695 6744 108 H(17A) 6943 −6807479 115 H(19A) 7126 −1628 8627 134 H(20A) 7766 −1689 7730 142 H(21A)5111 −624 9280 233 H(1A) 8376 −2045 5049 166 H(1B) 9947 −1923 5325 166U(eq) is defined as one third of the trace of the orthogonalized Uijtensor.

TABLE 5 Crystal Data of HCl salt: Form SA-1 Empirical formula C24 H26Cl3 N4 O2 Formula weight 508.84 Temperature 298(2) K Wavelength 1.54178A Crystal system, space group Monoclinic, P2₁ Unit cell dimensions a =11.0668(9) Å alpha = 90 deg. b = 7.3750(6) Å beta = 100.594(7) deg. c =15.3927(14) Å gamma = 90 deg. Volume 1234.90(18) Å³ Z, Calculateddensity 2, 1.363 Mg/m³ Absorption coefficient 3.595 mm⁻¹ F(000) 530Theta range for data collection 4.06 to 61.98 deg. Limiting indices −12<= h <= 12, −7 <= k <= 6, −17 <= l <= 15 Reflections collected/unique3911/2687 [R(int) = 0.0253] Completeness to theta = 61.98 89.5%Refinement method Full-matrix least-squares on F{circumflex over ( )}2Data/restraints/parameters 2687/1/306 Goodness-of-fit on F{circumflexover ( )}2 1.035 Final R indices [I > R1 = 0.0382, wR2 = 0.09942sigma(I)] R indices (all data) R1 = 0.0423, wR2 = 0.1027 Absolutestructure parameter 0.02(2) Largest diff. peak and hole 0.270 and −0.201e. Å⁻³

TABLE 6 Atomic Coordinates of HCl salt: Form SA-1 Atomic coordinates(×10⁴) and equivalent isotropic displacement parameters Å² × 10³) forForm SA-1. x y z U(eq) Cl 12265(1)  6142(1) 1683(1) 49(1) Cl(1) 7875(1)12955(2)  4765(1) 82(1) Cl(2) 8143(1) 9869(2) 6212(1) 87(1) N(1) 2603(2)8917(4) −585(2) 34(1) N(2) 10328(2)  9284(4) 1422(2) 39(1) C(3) 7992(3)8350(5) 1854(2) 31(1) C(4) 6974(3) 8951(5)  360(2) 32(1) N(5) 1421(3)9376(5) −494(2) 47(1) C(6) 5842(3) 8414(5)  549(2) 32(1) C(7) 4724(3)8458(5) −145(2) 32(1) C(8) 8036(3) 8902(5)  998(2) 31(1) C(9) 3613(3)8927(5)  63(2) 36(1) C(10) 9143(3) 8296(5) 2564(2) 35(1) N(11) 1476(3)8685(5) −1929(2)  51(1) C(12) 5807(3) 7820(6) 1405(2) 37(1) C(13)8878(3) 8695(5) 3475(2) 37(1) C(14) 6859(3) 7787(6) 2035(2) 38(1) C(15)4772(3) 8039(5) −1033(2)  41(1) C(16) 10107(3)  9607(5) 2333(2) 38(1)C(17) 2614(3) 8532(5) −1448(3)  39(1) C(18) 9221(3) 9458(6)  715(2)42(1) C(19) 8304(4) 10787(6)  4526(3) 47(1) C(20) 8550(3) 10430(5) 3699(3) 42(1) C(21) 3747(4) 8064(6) −1674(2)  46(1) C(22)  821(3)9193(6) −1314(3)  50(1) C(23) 8957(4) 7332(6) 4108(3) 48(1) C(24)8714(4) 7701(7) 4937(3) 55(1) C(25) 8399(4) 9426(8) 5162(3) 58(1) OW112197(4)  11835(6)  1559(3) 63(1) O(01) 13401(5)  9513(6) 2783(4)138(2)  C(01) 14893(7)   7959(17) 3801(5) 166(5)  C(02) 14430(8)  9598(14) 3370(6) 139(3)  C(03) 14517(9)  11360(20) 3818(8) 221(8) H(2A) 10639 8162 1397 46 H(2B) 10900 10076 1311 46 H(4A) 7017 9351 −20738 H(9A) 3554 9248 638 43 H(10A) 9484 7068 2573 42 H(12A) 5066 7445 154944 H(14A) 6817 7377 2600 46 H(15A) 5524 7738 −1183 49 H(16A) 9829 108442381 45 H(16B) 10871 9453 2750 45 H(18A) 9335 8717 216 50 H(18B) 914810709 518 50 H(20A) 8495 11359 3285 50 H(21A) 3795 7776 −2255 55 H(22A)−20 9407 −1461 60 H(23A) 9175 6163 3970 58 H(24A) 8763 6773 5351 66 HW112650(50)  11440(80) 1990(40)  67(19) HW2 12190(50)   12930(110)1710(40)  90(20) H(01D) 13362 8533 2528 207 H(01A) 14782 6981 3382 249H(01B) 14456 7696 4270 249 H(01C) 15752 8098 4041 249 H(02A) 15024 97772977 167 H(03A) 14198 12289 3401 331 H(03B) 15361 11617 4062 331 H(03C)14047 11331 4284 331 U(eq) is defined as one third of the trace of theorthogonalized Uij tensor.

TABLE 7 Crystal Data of HCl salt: Form N-2 Empirical formula C21 H17 Cl3N4 Formula weight 431.74 Temperature 298(2) K Wavelength 1.54178 ÅCrystal system, space group Orthorhombic, P2₁2₁2₁ Unit cell dimensions a= 7.1183(2) Å alpha = 90 deg. b = 21.2160(7) Å beta = 90 deg. c =26.3602(9) Å gamma = 90 deg. Volume 3981.0(2) Å³ Z, Calculated density8, 1.441 Mg/m³ Absorption coefficient 4.283 mm⁻¹ F(000) 1776 Crystalsize 0.16 × 0.07 × 0.06 mm Theta range for data collection 2.67 to 44.53deg. Limiting indices −6 <= h <= 5, −19 <= k <= 18, −23 <= l <= 23Reflections collected/unique 9626/2985 [R(int) = 0.0700] Completeness totheta = 44.53 95.3% Data/restraints/parameters 2985/0/505Goodness-of-fit on F{circumflex over ( )}2 1.031 Final R indices [I > R1= 0.0580, wR2 = 0.1446 2sigma(I)] R indices (all data) R1 = 0.0780, wR2= 0.1669 Absolute structure parameter 0.10(4) Largest diff. peak andhole 0.260 and −0.278 e. Å⁻³

TABLE 8 Atomic Coordinates of HCl salt: Form N-2 Atomic coordinates(×10⁴) and equivalent isotropic displacement parameters (Å² × 10³) forForm N-2. x y z U(eq) Cl(1) 4498(5)  2054(2) 5726(1) 84(1) Cl(2)8606(6)  2604(2) 5897(1) 98(1) Cl(3) 13423(5)  8143(1) 1794(1) 75(1)Cl(4) 9097(4)  8448(1) 1988(1) 73(1) Cl(5) −2074(4)  5119(1) 4228(1)71(1) Cl(6) 3031(4)  5078(1) 2983(1) 66(1) N(1) 2223(11) 4893(4) 4125(3)52(2) N(2)  61(15) 7409(6) 6214(5) 64(3) N(3) −573(13) 7985(6) 6078(5)65(3) N(4) −306(16) 7936(6) 6927(5) 75(4) N(5) 7228(10) 5382(4) 3091(3)47(2) N(6) 9780(14) 2724(5) 1073(5) 56(3) N(7) 10462(14)  2158(6)1235(4) 62(3) N(8) 10074(16)  2166(6)  367(4) 70(3) C(1) 3750(20)3157(6) 5294(4) 67(4) C(2) 5220(20) 2801(5) 5526(4) 62(4) C(3) 6990(20)3065(8) 5577(5) 75(4) C(4) 7330(20) 3646(7) 5390(5) 75(5) C(5) 5980(20)3987(6) 5149(5) 67(4) C(6) 4180(20) 3750(6) 5092(4) 57(4) C(7) 2634(17)4168(5) 4848(4) 53(3) C(8) 3267(15) 4321(5) 4307(4) 54(3) C(9) 2762(18)5465(5) 4424(5) 63(4) C(10) 2298(13) 5348(6) 4977(5) 44(3) C(11)2294(14) 4749(5) 5175(5) 42(3) C(12) 1796(17) 4667(5) 5682(5) 57(3)C(13) 1424(16) 5177(6) 5975(5) 57(3) C(14) 1510(15) 5791(5) 5785(5)45(3) C(15) 1928(14) 5865(5) 5284(5) 44(3) C(16) 1095(14) 6353(6)6107(5) 44(3) C(17)  466(16) 6920(7) 5908(5) 52(3) C(18) −747(19)8258(7) 6533(8) 79(5) C(19)  230(20) 7382(8) 6719(8) 79(4) C(20) 856(16) 6812(7) 6955(5) 61(3) C(21) 1241(15) 6307(6) 6639(6) 58(4)C(31) 11260(20)  6456(5) 2095(5) 68(4) C(32) 12471(16)  6939(6) 1978(4)63(4) C(33) 11878(19)  7564(6) 1953(4) 61(3) C(34) 9939(18) 7684(5)2033(4) 55(3) C(35) 8744(17) 7205(5) 2162(4) 51(3) C(36) 9370(18)6600(5) 2199(4) 52(3) C(37) 8002(17) 6074(5) 2356(4) 49(3) C(38)8399(14) 5938(5) 2920(4) 51(3) C(39) 7870(18) 4792(5) 2834(5) 60(4)C(40) 8081(17) 4873(6) 2263(5) 53(3) C(41) 8178(17) 5465(5) 2060(5)52(3) C(42) 8419(18) 5507(5) 1536(6) 66(4) C(43) 8611(16) 4964(7)1238(4) 59(3) C(44) 8532(16) 4370(6) 1459(5) 54(3) C(45) 8220(17)4337(5) 1978(5) 57(3) C(46) 8796(17) 3796(6) 1143(5) 54(3) C(47)9454(16) 3252(7) 1367(5) 56(3) C(48) 10601(16)  1851(6)  794(7) 67(4)C(49) 9511(17) 2725(6)  563(7) 55(4) C(50) 8909(16) 3292(7)  321(5)62(4) C(51) 8534(16) 3805(6)  614(6) 53(3) H(1A) 2481 4958 3795 62 H(1C)979 4827 4155 62 H(5A) 7327 5336 3429 56 H(5C) 6012 5453 3016 56 H(1B)2535 2999 5277 81 H(4B) 8526 3818 5427 90 H(5B) 6262 4384 5021 80 H(7B)1466 3924 4831 63 H(8B) 4609 4401 4302 65 H(8C) 3009 3966 4086 65 H(9A)2075 5829 4301 76 H(9B) 4095 5547 4386 76 H(12A) 1718 4264 5818 68H(13A) 1102 5116 6313 69 H(15A) 1967 6267 5145 52 H(17A) 322 6962 555962 H(18A) −1175 8671 6562 94 H(20A) 998 6783 7305 73 H(21A) 1607 59266783 70 H(31A) 11679 6042 2104 81 H(32A) 13726 6845 1914 76 H(35A) 74867294 2226 62 H(37A) 6713 6232 2322 59 H(38A) 9722 5846 2967 61 H(38B)8090 6306 3123 61 H(39A) 6970 4458 2901 71 H(39B) 9067 4664 2976 71H(42A) 8454 5901 1382 79 H(43A) 8793 5002 890 71 H(45A) 8104 3945 213369 H(47A) 9678 3241 1714 67 H(48A) 11041 1439 779 80 H(50A) 8777 3311−30 74 H(51A) 8094 4171 460 63 U(eq) is defined as one third of thetrace of the orthogonalized Uij tensor.

Example 11 Powder X-Ray Diffraction for Forms SA-1 and N-2

X-ray powder diffraction (PXRD) data were obtained using a Bruker C2GADDS. The radiation was Cu Kα (40 KV, 40 MA). The sample-detectordistance was 15 cm. Powder samples were placed in sealed glasscapillaries of 1 mm or less in diameter; the capillary was rotatedduring data collection. Data were collected for 3≦20≦35° with a sampleexposure time of at least 1000 seconds. The resulting two-dimensionaldiffraction arcs were integrated to create a traditional 1-dimensionalPXRD. The results of the PXRD pattern and a simulated pattern calculatedfrom the single crystal data for Form SA-1 are shown in FIG. 1.

Table 9 lists the characteristic PXRD peaks that describe Form SA-1((S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemonohydrochloride monoisopropanolate monohydrate) and Form N-2((S)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinolinemonohydrochloride). In particular, Table 9 shows characteristicdiffraction peak positions (degrees 20±0.1) at room temperature, basedon a high quality pattern collected with a diffractometer (cuKα) with aspinning capillary with 20 calibrated with a NIST or other suitablestandard.

TABLE 9 Form SA-1 Form N-2 5.8 8.3 8.1 8.9 9.1 10.9 10.8 14.2 11.7 14.713.0 16.7 13.3 17.3 14.5 18.0 15.1 18.4 15.4 18.8 16.2 20.2 16.8 21.9

Example 12 Differential Scanning Calorimetry for Form SA-1

Differential scanning calorimetry (DSC) experiments were performed in aTA Instruments™ model Q1000 or 2920. The sample (about 2-6 mg) wasweighed in a pinpricked hermetically sealed aluminum pan and accuratelyrecorded to a hundredth of a milligram, and transferred to the DSC. Theinstrument was purged with nitrogen gas at 50 mL/min. Data werecollected between room temperature and 300° C. at 10° C.min. heatingrate. The plot was made with the endothermic peaks pointing down. Theresults are shown in FIG. 2.

Example 13 Thermogravimetric Analysis for Form SA-1

The results are shown in FIG. 3.

Although preferred embodiments have been depicted and described indetail herein, it will be apparent to those skilled in the relevant artthat various modifications, additions, substitutions, and the like canbe made without departing from the spirit of the invention and these aretherefore considered to be within the scope of the invention as definedin the claims which follow.

1. Crystalline Form SA-1 of

wherein: the carbon atom designated * is in the S configuration; or a pharmaceutically acceptable salt thereof or a solvate thereof.
 2. Crystalline Form SA-1 of

wherein: the carbon atom designated * is in the S configuration, characterized by the following unit cell parameters: Cell dimensions: a=11.0668(9) Å b=7.3750(6) Å c=15.3927(14) Å alpha=90° beta=100.594(7)° gamma =90° Space group: Monoclinic, P2₁ Volume: 1234.90(18) Å³ Z, Calculated Density: 2, 1.363 Mg/m³, wherein measurement of said crystalline form is at a temperature of between about 20° C. to about 25° C.
 3. Crystalline Form SA-1 of

wherein: the carbon atom designated * is in the S configuration, characterized by fractional atomic coordinates within the unit cell as listed in Table
 6. 4. Crystalline Form SA-1 of

wherein: the carbon atom designated * is in the S configuration, with characteristic peaks in a powder X-ray diffraction pattern at values of 2 theta of 5.8±0.1, 8.1±0.1, 9.1±0.1, 10.8±0.1, 11.7±0.1, 13.0±0.1, 13.3±0.1, 14.5±0.1, 15.1±0.1, 15.4±0.1, 16.2±0.1, and 16.8±0.1, at a temperature between about 20° C. and about 25° C.
 5. Crystalline Form SA-1 of

wherein: the carbon atom designated * is in the S configuration, characterized by a melt with decomposition endotherm with onset of about 85° C.
 6. Substantially pure crystalline Form SA-1 of

wherein: the carbon atom designated * is in the S configuration.
 7. The crystalline form of claim 6, wherein said Form SA-1Has a purity of at least 98 wt %.
 8. The crystalline form of claim 6, wherein said Form SA-1Has a purity of at least 99 wt %.
 9. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of the crystalline form according to claim
 1. 10. A method of treating a disorder which is created by or is dependent upon decreased availability of norepinephrine, dopamine, or serotonin, said method comprising: administering to a patient in need of such treatment a therapeutically effective amount of a crystalline form according to claim 1 or a pharmaceutically acceptable salt thereof.
 11. The method according to claim 10, wherein the disorder is selected from the group consisting of attention deficit hyperactivity disorder (ADHD), cognition impairment, anxiety disorders, generalized anxiety disorder (GAD), panic disorder, bipolar disorder or manic depression or manic-depressive disorder, obsessive compulsive disorder (OCD), posttraumatic stress disorder (PTSD), acute stress disorder, social phobia, simple phobias, pre-menstrual dysphoric disorder (PMDD), social anxiety disorder (SAD), major depressive disorder (MDD), postnatal depression, dysthymia, depression associated with Alzheimer's disease, Parkinson's disease, or psychosis, supranuclear palsy, eating disorders, obesity, anorexia nervosa, bulimia nervosa, binge eating disorder, diabetes, ischemic diseases, pain, substance abuse disorders, chemical dependencies, nicotine addiction, cocaine addiction, amphetamine addiction, alcohol addiction, Lesch-Nyhan syndrome, neurodegenerative diseases, Parkinson's disease, late luteal phase syndrome or narcolepsy, psychiatric symptoms, anger, rejection sensitivity, movement disorders, extrapyramidal syndrome, Tic disorders, restless leg syndrome (RLS), tardive dyskinesia, supranuclear palsy, sleep related eating disorder (SRED), night eating syndrome (NES), stress urinary incontinence (SUI), migraine, neuropathic pain, diabetic neuropathy, lower back pain, fibromyalgia syndrome (FS), osteoarthritis pain, arthritis pain, chronic fatigue syndrome (CFS), sexual dysfunction, premature ejaculation, male impotence, thermoregulatory disorders (e.g., hot flashes associated with menopause), and irritable bowel syndrome (IBS).
 12. The method according to claim 10 further comprising: administering a therapeutically effective amount of a serotonin 1A receptor antagonist or a pharmaceutically acceptable salt thereof.
 13. The method according to claim 12, wherein the serotonin 1A receptor antagonist is selected from the group consisting of WAY 100135 and spiperone.
 14. The method according to claim 10 further comprising: administering a therapeutically effective amount of a selective neurokinin-1 receptor antagonist or a pharmaceutically acceptable salt thereof.
 15. The method according to claim 10 further comprising: administering a therapeutically effective amount of a norepinephrine precursor or a pharmaceutically acceptable salt thereof.
 16. The method according to claim 15, wherein the norepinephrine precursor is selected from the group consisting of L-tyrosine and L-phenylalanine.
 17. A method of inhibiting synaptic norepinephrine uptake in a patient comprising: administering to the patient a therapeutically effective inhibitory amount of a crystalline form according to claim 1 or a pharmaceutically acceptable salt thereof.
 18. A method of inhibiting synaptic serotonin uptake in a patient comprising: administering to the patient a therapeutically effective inhibitory amount of a crystalline form according to claim 1 or a pharmaceutically acceptable salt thereof.
 19. A method of inhibiting synaptic dopamine uptake in a patient comprsing: administering to the patient a therapeutically effective inhibitory amount of a crystalline form according to claim 1 or a pharmaceutically acceptable salt thereof.
 20. A method of suppressing the desire of humans to smoke comprising: administering to a human in need of such suppression an effective amount, to relieve the desire to smoke, of a crystalline form according to claim 1 or a pharmaceutically acceptable salt thereof.
 21. A method of suppressing the desire of humans to consume alcohol comprising: administering to a human in need of such suppression an effective amount, to relieve the desire to consume alcohol, of a crystalline form according to claim 1 or a pharmaceutically acceptable salt thereof.
 22. Crystalline Form N-2 of

wherein: the carbon atom designated * is in the S configuration; or a pharmaceutically acceptable salt thereof or a solvate thereof.
 23. Crystalline Form N-2 of

wherein: the carbon atom designated * is in the S configuration, characterized by the following unit cell parameters: Cell dimensions: a=7.1183(2) Å b=21.2160(7) Å c=26.3602(9) Å alpha=90° beta=90° gamma=90° Space group: Orthorhombic, P2₁2₁2₁ Volume: 3981.0(2) Å³ Z, Calculated Density: 8, 1.441 Mg/m³ wherein measurement of said crystalline form is at a temperature of between about 20° C. to about 25° C.
 24. Crystalline Form N-2 of

wherein: the carbon atom designated * is in the S configuration, characterized by fractional atomic coordinates within the unit cell as listed in Table
 8. 25. Crystalline Form N-2 of

wherein: the carbon atom designated * is in the S configuration, with characteristic peaks in a powder X-ray diffraction pattern at values of 2 theta of 8.3±0.1, 8.9±0.1, 10.9±0.1, 14.2±0.1, 14.7±0.1, 16.7±0.1, 17.3±0.1, 18.0±0.1, 18.4±0.1, 18.8±0.1, 20.2±0.1, and 21.9±0.1, and 16.8±0.1, at a temperature between about 20° C. and about 25° C.
 26. Crystalline Form N-2 of

wherein: the carbon atom designated * is in the S configuration, characterized by a melt with decomposition endotherm with onset of about 250° C.
 27. Substantially pure crystalline Form N-2 of

wherein: the carbon atom designated * is in the S configuration.
 28. The crystalline form of claim 27, wherein said Form N-2Has a purity of at least 98 wt %.
 29. The crystalline form of claim 27, wherein said Form N-2Has a purity of at least 99 wt %.
 30. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of the crystalline form according to claim
 22. 31. A method of treating a disorder which is created by or is dependent upon decreased availability of norepinephrine, dopamine, or serotonin, said method comprising: administering to a patient in need of such treatment a therapeutically effective amount of a crystalline form according to claim 22 or a pharmaceutically acceptable salt thereof.
 32. The method according to claim 31, wherein the disorder is selected from the group consisting of attention deficit hyperactivity disorder (ADHD), cognition impairment, anxiety disorders, generalized anxiety disorder (GAD), panic disorder, bipolar disorder or manic depression or manic-depressive disorder, obsessive compulsive disorder (OCD), posttraumatic stress disorder (PTSD), acute stress disorder, social phobia, simple phobias, pre-menstrual dysphoric disorder (PMDD), social anxiety disorder (SAD), major depressive disorder (MDD), postnatal depression, dysthymia, depression associated with Alzheimer's disease, Parkinson's disease, or psychosis, supranuclear palsy, eating disorders, obesity, anorexia nervosa, bulimia nervosa, binge eating disorder, diabetes, ischemic diseases, pain, substance abuse disorders, chemical dependencies, nicotine addiction, cocaine addiction, amphetamine addiction, alcohol addiction, Lesch-Nyhan syndrome, neurodegenerative diseases, Parkinson's disease, late luteal phase syndrome or narcolepsy, psychiatric symptoms, anger, rejection sensitivity, movement disorders, extrapyramidal syndrome, Tic disorders, restless leg syndrome (RLS), tardive dyskinesia, supranuclear palsy, sleep related eating disorder (SRED), night eating syndrome (NES), stress urinary incontinence (SUI), migraine, neuropathic pain, diabetic neuropathy, lower back pain, fibromyalgia syndrome (FS), osteoarthritis pain, arthritis pain, chronic fatigue syndrome (CFS), sexual dysfunction, premature ejaculation, male impotence, thermoregulatory disorders (e.g., hot flashes associated with menopause), and irritable bowel syndrome (IBS).
 33. The method according to claim 31 further comprising: administering a therapeutically effective amount of a serotonin 1A receptor antagonist or a pharmaceutically acceptable salt thereof.
 34. The method according to claim 33, wherein the serotonin 1A receptor antagonist is selected from the group consisting of WAY 100135 and spiperone.
 35. The method according to claim 31 further comprising: administering a therapeutically effective amount of a selective neurokinin-1 receptor antagonist or a pharmaceutically acceptable salt thereof.
 36. The method according to claim 31 further comprising: administering a therapeutically effective amount of a norepinephrine precursor or a pharmaceutically acceptable salt thereof.
 37. The method according to claim 36, wherein the norepinephrine precursor is selected from the group consisting of L-tyrosine and L-phenylalanine.
 38. A method of inhibiting synaptic norepinephrine uptake in a patient comprising: administering to the patient a therapeutically effective inhibitory amount of a crystalline form according to claim 22 or a pharmaceutically acceptable salt thereof.
 39. A method of inhibiting synaptic serotonin uptake in a patient comprising: administering to the patient a therapeutically effective inhibitory amount of a crystalline form according to claim 22 or a pharmaceutically acceptable salt thereof.
 40. A method of inhibiting synaptic dopamine uptake in a patient comprsing: administering to the patient a therapeutically effective inhibitory amount of a crystalline form according to claim 22 or a pharmaceutically acceptable salt thereof.
 41. A method of suppressing the desire of humans to smoke comprising: administering to a human in need of such suppression an effective amount, to relieve the desire to smoke, of a crystalline form according to claim 22 or a pharmaceutically acceptable salt thereof.
 42. A method of suppressing the desire of humans to consume alcohol comprising: administering to a human in need of such suppression an effective amount, to relieve the desire to consume alcohol, of a crystalline form according to claim 22 or a pharmaceutically acceptable salt thereof.
 43. A process for preparation of a product compound of Formula (I)

wherein the carbon atom designated * is in the R or S configuration, said process comprising: treating a first intermediate compound of Formula (II):

with an acid under conditions effective to produce the product compound.
 44. The process according to claim 43 wherein the acid is selected from the group consisting of sulfuric acid, methansulfonic acid, phosphoric acid, and L-tartaric acid. 